Compositions for immunising against staphylococcus aureus

ABSTRACT

An effective  Staphylococcus aureus  vaccine may require several antigenic components, and so various combinations of  S. aureus  antigens are identified for use in immunisation. These polypeptides may optionally be used in combination with  S. aureus  saccharides.

TECHNICAL FIELD

This invention relates to antigens derived from S. aureus and to theiruse in immunisation.

BACKGROUND ART

Staphylococcus aureus is a Gram-positive spherical bacterium. Annual USmortality exceeds that of any other infectious disease, includingHIV/AIDS, and S. aureus is the leading cause of bloodstream, lowerrespiratory tract, skin & soft tissue infections. There is currently noauthorised vaccine. A vaccine based on a mixture of surfacepolysaccharides from bacterial types 5 and 8, StaphVAX™, failed toreduce infections when compared to the placebo group in a phase IIIclinical trial in 2005.

Reference 1 reports that the “V710” vaccine from Merck and Intercell isundergoing a phase 2/3 trial on patients undergoing cardiothoracicsurgery. The V710 vaccine is based on a single antigen, IsdB [2], aconserved iron-sequestering cell-surface protein.

S. aureus causes a range of illnesses from minor skin infections tolife-threatening diseases such as pneumonia, meningitis, osteomyelitis,bacteremia, endocarditis, toxic shock syndrome, organ abscesses andsepticemia. The bacterium has multiple virulence factors which aredifferentially expressed during different phases of its life cycle, andso a vaccine which can prevent one disease might not prevent another.For instance, the V710 vaccine may be effective against hematic spreadof the S. aureus, but may be ineffective against pneumonia and may notelicit any opsonic activity. One aim of the invention is to providevaccines which can protect against hematic spread and pneumonia, andwhich may also elicit an opsonic response.

Thus there remains a need to identify further and improved antigens foruse in S. aureus vaccines, and in particular for vaccines which areuseful against multiple S. aureus pathologies.

DISCLOSURE OF THE INVENTION

The inventors have identified various S. aureus polypeptides that areuseful for immunisation, either alone or in combination. Thesepolypeptides may be combined with S. aureus saccharides or other S.aureus polypeptides. The antigens are useful in S. aureus vaccines butmay also be used as components in vaccines for immunising againstmultiple pathogens.

The inventors have identified the following 36 polypeptides: clfA, clfB,coA, eap, ebhA, ebpS, efb, emp, esaC, esxA, esxB, FnBA, FnBB, Hla, hlgB,hlgC, isdA, isdB, isdC, isdG, isdH, isdI, lukD, lukE, lukF, lukS, nuc,sasA, sasB, sasC, sasD, sasF, sdrC, sdrD, spa, and sdrE2. This set ofantigens is referred to herein as ‘the first antigen group’. Thus theinvention provides an immunogenic composition comprising a combinationof antigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6or more) antigens selected from the group consisting of: (1) a clfAantigen; (2) a clfB antigen; (3) a coA antigen; (4) a eap antigen; (5) aebhA antigen; (6) a ebpS antigen; (7) a efb antigen; (8) a emp antigen;(9) a esaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) aFnBA antigen; (13) a FnBB antigen; (14) a Hla antigen; (15) a hlgBantigen; (16) a hlgC antigen; (17) a isdA antigen; (18) a isdB antigen;(19) a isdC antigen; (20) a isdG antigen; (21) a isdH antigen; (22) aisdI antigen; (23) a lukD antigen; (24) a lukE antigen; (25) a lukFantigen; (26) a lukS antigen; (27) a nuc antigen; (28) a sasA antigen;(29) a sasB antigen; (30) a sasC antigen; (31) a sasD antigen; (32) asasF antigen; (33) a sdrC antigen; (34) a sdrD antigen; (35) a spaantigen; (36) a sdrE2 antigen.

Within the first antigen group, antigens are preferably selected from asubset of 16 of the 36 polypeptides, namely: clfA, clfB, emp, esaC,esxA, esxB, hla, isdA, isdB, isdC, sasD, sasF, sdrC, sdrD, spa, andsdrE2. Thus the invention provides an immunogenic composition comprisinga combination of antigens, said combination comprising two or more (i.e.2, 3, 4, 5, 6 or more) antigens selected from the group consisting ofthese sixteen antigens.

The inventors have also identified the following 128 polypeptides:sta001, sta002, sta003, sta004, sta005, sta006, sta007, sta008, sta009,sta010, sta011, sta012, sta013, sta014, sta015, sta016, sta017, sta018,sta019, sta020, sta021, sta022, sta023, sta024, sta025, sta026, sta027,sta028, sta029, sta030, sta031, sta032, sta033, sta034, sta035, sta036,sta037, sta038, sta039, sta040, sta041, sta042, sta043, sta044, sta045,sta046, sta047, sta048, sta049, sta050, sta051, sta052, sta053, sta054,sta055, sta056, sta057, sta058, sta059, sta060, sta061, sta062, sta063,sta064, sta065, sta066, sta067, sta068, sta069, sta070, sta071, sta072,sta073, sta074, sta075, sta076, sta077, sta078, sta079, sta080, sta081,sta082, sta083, sta084, sta085, sta086, sta087, sta088, sta089, sta090,sta091, sta092, sta093, sta094, sta095, sta096, sta097, sta098, sta099,sta100, sta101, sta102, sta103, sta104, sta105, sta106, sta107, sta108,sta109, sta110, sta111, sta112, sta113, sta114, sta115, sta116, sta117,sta118, sta119, sta120, NW_(—)6, NW_(—)9, NW_(—)10, NW_(—)7, NW_(—)8,NW_(—)2, NW_(—)1, and NW_(—)5. This set of antigens is referred toherein as ‘the second antigen group’. Thus the invention provides animmunogenic composition comprising a combination of antigens, saidcombination comprising two or more (i.e. 2, 3, 4, 5, 6 or more) antigensselected from the group consisting of: (1) a sta001 antigen; (2) asta002 antigen; (3) a sta003 antigen; (4) a sta004 antigen; (5) a sta005antigen; (6) a sta006 antigen; (7) a sta007 antigen; (8) a sta008antigen; (9) a sta009 antigen; (10) a sta010 antigen; (11) a sta011antigen; (12) a sta012 antigen; (13) a sta013 antigen; (14) a sta014antigen; (15) a sta015 antigen; (16) a sta016 antigen; (17) a sta017antigen; (18) a sta018 antigen; (19) a sta019 antigen; (20) a sta020antigen; (21) a sta021 antigen; (22) a sta022 antigen; (23) a sta023antigen; (24) a sta024 antigen; (25) a sta025 antigen; (26) a sta026antigen; (27) a sta027 antigen; (28) a sta028 antigen; (29) a sta029antigen; (30) a sta030 antigen; (31) a sta031 antigen; (32) a sta032antigen; (33) a sta033 antigen; (34) a sta034 antigen; (35) a sta035antigen; (36) a sta036 antigen; (37) a sta037 antigen; (38) a sta038antigen; (39) a sta039 antigen; (40) a sta040 antigen; (41) a sta041antigen; (42) a sta042 antigen; (43) a sta043 antigen; (44) a sta044antigen; (45) a sta045 antigen; (46) a sta046 antigen; (47) a sta047antigen; (48) a sta048 antigen; (49) a sta049 antigen; (50) a sta050antigen; (51) a sta051 antigen; (52) a sta052 antigen; (53) a sta053antigen; (54) a sta054 antigen; (55) a sta055 antigen; (56) a sta056antigen; (57) a sta057 antigen; (58) a sta058 antigen; (59) a sta059antigen; (60) a sta060 antigen; (61) a sta061 antigen; (62) a sta062antigen; (63) a sta063 antigen; (64) a sta064 antigen; (65) a sta065antigen; (66) a sta066 antigen; (67) a sta067 antigen; (68) a sta068antigen; (69) a sta069 antigen; (70) a sta070 antigen; (71) a sta071antigen; (72) a sta072 antigen; (73) a sta073 antigen; (74) a sta074antigen; (75) a sta075 antigen; (76) a sta076 antigen; (77) a sta077antigen; (78) a sta078 antigen; (79) a sta079 antigen; (80) a sta080antigen; (81) a sta081 antigen; (82) a sta082 antigen; (83) a sta083antigen; (84) a sta084 antigen; (85) a sta085 antigen; (86) a sta086antigen; (87) a sta087 antigen; (88) a sta088 antigen; (89) a sta089antigen; (90) a sta090 antigen; (91) a sta091 antigen; (92) a sta092antigen; (93) a sta093 antigen; (94) a sta094 antigen; (95) a sta095antigen; (96) a sta096 antigen; (97) a sta097 antigen; (98) a sta098antigen; (99) a sta099 antigen; (100) a sta100 antigen; (101) a sta101antigen; (102) a sta102 antigen; (103) a sta103 antigen; (104) a sta104antigen; (105) a sta105 antigen; (106) a sta106 antigen; (107) a sta107antigen; (108) a sta108 antigen; (109) a sta109 antigen; (110) a sta110antigen; (111) a sta111 antigen; (112) a sta12 antigen; (113) a sta113antigen; (114) a sta114 antigen; (115) a sta115 antigen; (116) a sta116antigen; (117) a sta117 antigen; (118) a sta118 antigen; (119) a sta119antigen; (120) a sta120 antigen; (121) a NW_(—)6 antigen; (122) aNW_(—)9 antigen; (123) a NW_(—)10 antigen; (124) a NW_(—)7 antigen;(125) a NW_(—)8 antigen; (126) a NW_(—)2 antigen; (127) a NW_(—)1antigen; and (128) a NW_(—)5 antigen.

Within the second antigen group of 128 antigens, a preferred subset of113 antigens omits (81) and (107) to (120) from this list.

Within the second antigen group, a subset of 27 of the 128 polypeptidesis referred to herein as ‘the third antigen group’, namely: sta001,sta002, sta003, sta004, sta005, sta006, sta007, sta008, sta009, sta010,sta019, sta028, sta040, sta049, sta057, sta064, sta073, sta095, sta098,sta101, sta105, NW_(—)1, NW_(—)6, NW_(—)7, NW_(—)8, NW_(—)9 andNW_(—)10. The invention provides an immunogenic composition comprising acombination of antigens, said combination comprising two or more (i.e.2, 3, 4, 5, 6 or more) antigens selected from the third antigen group.

The 101 antigens that are in the second antigen group but not in thethird antigen group are referred to herein as ‘the fourth antigengroup’. Within the fourth antigen group of 101 antigens, a preferredsubset of 86 antigens omits (81) and (107) to (120) from the above list.The second antigen group thus consists of a combination of the third andfourth antigen groups.

Within the second antigen group, a subset of 8 of the 128 polypeptidesis referred to herein as ‘the fifth antigen group’, namely: sta004,sta006, sta007, sta011, sta028, sta060, sta098 and sta112. The inventionprovides an immunogenic composition comprising a combination ofantigens, said combination comprising two or more (i.e. 2, 3, 4, 5, 6 ormore) antigens selected from the fifth antigen group.

Within the 36 antigens of the first antigen group there are 630 possiblepairs of different antigens. All such pairs are disclosed herein and arepart of the invention. Thus the invention provides an immunogeniccomposition comprising a pair of antigens, wherein said pair is one ofsaid 630 pairs.

Within the 128 antigens of the second antigen group there are 8128possible pairs of different antigens. All such pairs are disclosedherein and are part of the invention. Thus the invention provides animmunogenic composition comprising a pair of antigens, wherein said pairis one of said 8128 pairs.

Within the preferred 113 antigens of the second antigen group there are6328 possible pairs of different antigens. All such pairs are disclosedherein and are part of the invention. Thus the invention provides animmunogenic composition comprising a pair of antigens, wherein said pairis one of said 6328 pairs.

Within the preferred 27 antigens of the third antigen group there are351 possible pairs of different antigens. All such pairs are disclosedherein and are part of the invention. Thus the invention provides animmunogenic composition comprising a pair of antigens, wherein said pairis one of said 351 pairs.

Within the 101 antigens of the fourth antigen group there are 5050possible pairs of different antigens. All such pairs are disclosedherein and are part of the invention. Thus the invention provides animmunogenic composition comprising a pair of antigens, wherein said pairis one of said 5050 pairs.

Within the preferred 86 antigens of the fourth antigen group there are3655 possible pairs of different antigens. All such pairs are disclosedherein and are part of the invention. Thus the invention provides animmunogenic composition comprising a pair of antigens, wherein said pairis one of said 3655 pairs.

In one embodiment, a composition includes at least one antigen (i.e. 1,2, 3, 4, 5, 6 or more) selected from the first antigen group and atleast one antigen (i.e. 1, 2, 3, 4, 5, 6 or more) selected from thesecond antigen group. Antigens from the first antigen group may beselected from the preferred subset of 16 antigens, and antigens from thesecond antigen group may be selected from the third antigen group or thefifth antigen group.

The invention also provides an immunogenic composition comprising acombination of antigens, said combination comprising two or more (i.e.2, 3, 4, 5, 6 or more) antigens selected from the group consisting of:(1) a clfA antigen; (2) a clfB antigen; (3) a sdrE2 antigen; (4) a sdrCantigen; (5) a SasF antigen; (6) a emp antigen; (7) a sdrD antigen; (8)a spa antigen; (9) a esaC antigen; (10) a esxA antigen; (11) a esxBantigen; (12) a sta006 antigen; (13) a isdC antigen; (14) a Ma antigen;(15) a sta011 antigen; (16) isdA antigen; (17) a isdB antigen; (18) asasF antigen. This group of 18 antigens is sometimes referred to hereinas the ‘sixth antigen group’.

The invention also provides an immunogenic composition comprising acombination of antigens, said combination comprising two or more (i.e.2, 3, 4 or 5) antigens selected from the group consisting of: (1) a esxAantigen; (2) a esxB antigen; (3) a sta006 antigen; (4) a hla antigen;and/or (5) a sta011 antigen. The composition may also include anadjuvant e.g. an aluminium hydroxide adjuvant.

Advantageous combinations of the invention are those in which two ormore antigens act synergistically. Thus the protection against S. aureusdisease achieved by their combined administration exceeds that expectedby mere addition of their individual protective efficacy.

Specific combinations of interest include, but are not limited to:

(1) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen and a isdC antigen. The sdrD and sdrE2 antigens can usefully becombined as a hybrid polypeptide, as discussed below, e.g. an SdrDEhybrid with an sdrE2 antigen downstream of a sdrD antigen.(2) An immunogenic composition comprising a sasD antigen, a clfB antigenand a sdrC antigen.(3) An immunogenic composition comprising a sasD antigen, a clfBantigen, a sdrC antigen and a clfA antigen.(4) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen, a isdC antigen and a sta011 antigen. The sdrD and sdrE2antigens can usefully be combined as a hybrid polypeptide, as discussedbelow, e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrDantigen.(5) An immunogenic composition comprising a sasD antigen, a clfBantigen, a sdrC antigen and a sta006 antigen.(6) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen, a isdC antigen and a hla antigen. The sdrD and sdrE2 antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.The Hla antigen may be a detoxified mutant e.g. including a H35Lmutation.(7) An immunogenic composition comprising a sasD antigen, a clfBantigen, a sdrC antigen and a esxA antigen.(8) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen and a hla antigen. The esxA and esxB antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.The Hla antigen may be a detoxified mutant e.g. including a H35Lmutation.(9) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen, a isdC antigen and a esxA antigen. The sdrD and sdrE2 antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.(10) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen and a sta011 antigen. The esxA and esxBantigens may be combined as a hybrid polypeptide, as discussed below,e.g. an EsxAB hybrid.(11) An immunogenic composition comprising a esxA antigen, a esxBantigen and a sta011 antigen. The esxA and esxB antigens can usefully becombined as a hybrid polypeptide, as discussed below, e.g. a EsxABhybrid with a esxB antigen downstream of a esxA antigen.(12) An immunogenic composition comprising a sasD antigen, a clfBantigen, a sdrC antigen and a spa antigen.(13) An immunogenic composition comprising a esxA antigen, a esxBantigen, a isdA antigen, a sta006 antigen, a sta011 antigen and a spaantigen. The esxA and esxB antigens may be combined as a hybridpolypeptide, as discussed below, e.g. an EsxAB hybrid. The isdA antigenmay be a fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. Thespa antigen may be a fragment of a full-length spa antigen, such as aSpa(D) domain mutated to disrupt or decrease binding to IgG Fc.(14) An immunogenic composition comprising a esxA antigen, a esxBantigen, a Hla antigen, a sta006 antigen and a sta011 antigen. The esxAand esxB antigens may be combined as a hybrid polypeptide, as discussedbelow, e.g. an EsxAB hybrid. The Hla antigen may be a detoxified mutante.g. including a H35L mutation.(15) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen, a isdC antigen and a sdrE2 antigen. The sdrD and sdrE2 antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.(16) An immunogenic composition comprising a esxA antigen, a esxBantigen and a hla antigen. The esxA and esxB antigens can usefully becombined as a hybrid polypeptide, as discussed below, e.g. a EsxABhybrid with a esxB antigen downstream of a esxA antigen. The Hla antigenmay be a detoxified mutant e.g. including a H35L mutation.(17) An immunogenic composition comprising a hla antigen, a isdAantigen, a sta006 antigen and a sta011 antigen. The isdA antigen may bea fragment of a full-length isdA antigen e.g. SEQ ID NO: 157. The Hlaantigen may be a detoxified mutant e.g. including a H35L mutation.(18) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen and a isdA antigen. The esxA and esxB antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a EsxAB hybrid with a esxB antigen downstream of a esxA antigen.The isdA antigen may be a fragment of a full-length isdA antigen e.g.SEQ ID NO: 157.(19) An immunogenic composition comprising a sasD antigen, a clfBantigen, a sdrC antigen and a hla antigen. The Hla antigen may be adetoxified mutant e.g. including a H35L mutation.(20) An immunogenic composition comprising a Hla antigen, a sta006antigen and a sta011 antigen. The Hla antigen may be a detoxified mutante.g. including a H35L mutation.(21) An immunogenic composition comprising a esxA antigen and a esxBantigen. The esxA and esxB antigens can usefully be combined as a hybridpolypeptide, as discussed below, e.g. an EsxAB hybrid with an esxBantigen downstream of an esxA antigen.(22) An immunogenic composition comprising a esxA antigen, a esxBantigen and a sta006 antigen. The esxA and esxB antigens can usefully becombined as a hybrid polypeptide, as discussed below, e.g. a EsxABhybrid with a esxB antigen downstream of a esxA antigen.(23) An immunogenic composition comprising a spa antigen, a sta006antigen and a sta011 antigen. The spa antigen may be a fragment of afull-length spa antigen, such as a Spa(D) domain mutated to disrupt ordecrease binding to IgG Fc.(24) An immunogenic composition comprising a esxA antigen, a esxBantigen, a isdA antigen, a sta006 antigen and a sta011 antigen. The esxAand esxB antigens may be combined as a hybrid polypeptide, as discussedbelow, e.g. an EsxAB hybrid. The isdA antigen may be a fragment of afull-length isdA antigen e.g. SEQ ID NO: 157.(25) An immunogenic composition comprising a sta006 antigen and a sta011antigen.(26) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen, a isdA antigen and a clfB antigen. The esxAand esxB antigens can usefully be combined as a hybrid polypeptide, asdiscussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of aesxA antigen. The isdA antigen may be a fragment of a full-length isdAantigen e.g. SEQ ID NO: 157. The clfB antigen may be a fragment of afull-length clfB antigen e.g. SEQ ID NO: 163.(27) An immunogenic composition comprising a sta006 antigen, a sta011antigen and a sta019 antigen.(28) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen, a hla antigen and a clfB antigen. The esxAand esxB antigens can usefully be combined as a hybrid polypeptide, asdiscussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of aesxA antigen. The clfB antigen may be a fragment of a full-length clfBantigen e.g. SEQ ID NO: 163. The Hla antigen may be a detoxified mutante.g. including a H35L mutation.(29) An immunogenic composition comprising a sta006 antigen, a sta011antigen, a sta019 antigen, and a hla antigen. The Hla antigen may be adetoxified mutant e.g. including a H35L mutation.(30) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen, a sta011 antigen and a clfB antigen. The esxAand esxB antigens can usefully be combined as a hybrid polypeptide, asdiscussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of aesxA antigen. The elm antigen may be a fragment of a full-length elmantigen e.g. SEQ ID NO: 163.(31) An immunogenic composition comprising a spa antigen, a esxAantigen, a esxB antigen, a sta006 antigen and a sta011 antigen. The spaantigen may be a fragment of a full-length spa antigen, such as a Spa(D)domain mutated to disrupt or decrease binding to IgG Fc. The esxA andesxB antigens may be combined as a hybrid polypeptide, as discussedbelow, e.g. an EsxAB hybrid.(32) An immunogenic composition comprising a sdrD antigen, a sdrE2antigen, a isdC antigen and a esxB antigen. The sdrD and sdrE2 antigenscan usefully be combined as a hybrid polypeptide, as discussed below,e.g. a SdrDE hybrid with a sdrE2 antigen downstream of a sdrD antigen.(33) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen, a sta011 antigen and a sta019 antigen. TheesxA and esxB antigens can usefully be combined as a hybrid polypeptide,as discussed below, e.g. a EsxAB hybrid with a esxB antigen downstreamof a esxA antigen.(34) An immunogenic composition comprising a esxA antigen, a esxBantigen, a sta006 antigen, a isdA antigen and a sdrD antigen. The esxAand esxB antigens can usefully be combined as a hybrid polypeptide, asdiscussed below, e.g. a EsxAB hybrid with a esxB antigen downstream of aesxA antigen. The isdA antigen may be a fragment of a full-length isdAantigen e.g. SEQ ID NO: 157. The sdrD antigen may be a fragment of afull-length sdrD antigen e.g. SEQ ID NO: 156.(35) An immunogenic composition comprising a esxA antigen, a esxBantigen, and a isdA antigen. The esxA and esxB antigens can usefully becombined as a hybrid polypeptide, as discussed below, e.g. a EsxABhybrid with a esxB antigen downstream of a esxA antigen. The isdAantigen may be a fragment of a full-length isdA antigen e.g. SEQ ID NO:157.(36) An immunogenic composition comprising a sasD antigen, a elmantigen, a sdrC antigen, a esxA antigen and a esxB antigen. The esxA andesxB antigens can usefully be combined as a hybrid polypeptide, asdiscussed below, e.g. an EsxAB hybrid with an esxB antigen downstream ofan esxA antigen.(37) An immunogenic composition comprising a Hla antigen, a spa antigen,a sta006 antigen and a sta011 antigen. The Hla antigen may be adetoxified mutant e.g. including a H35L mutation. The spa antigen may bea fragment of a full-length spa antigen, such as a Spa(D) domain mutatedto disrupt or decrease binding to IgG Fc.

In some embodiments, any of these 37 compositions may include additionalstaphylococcal antigens, and these further antigens can be polypeptidesand/or saccharides. For example, they can usefully also include one ormore S. aureus capsular saccharide conjugate(s) e.g. against a serotype5 and/or a serotype 8 strain. The inclusion of one or both suchconjugates is particularly useful for combinations (8), (10), (20),(23), (25), (31) and (37).

In other embodiments, these 37 compositions include no additionalstaphylococcal polypeptide antigens. In other embodiments, these 37compositions include no additional staphylococcal antigens. In otherembodiments, these 37 compositions include no additional antigens.

The invention also provides a polypeptide comprising amino acid sequence(a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 151; and/or (b)comprising a fragment of at least ‘n’ consecutive amino acids from aminoacids 1-97 of SEQ ID NO: 151 and at least ‘n’ consecutive amino acidsfrom amino acids 104-207 of SEQ ID NO: 151, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). The invention also provides a polypeptidecomprising amino acid sequence (a) having 80% or more identity (e.g.80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 152; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids from amino acids 1-104 of SEQ ID NO: 152 andat least ‘n’ consecutive amino acids from amino acids 111-207 of SEQ IDNO: 152, wherein ‘n’ is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25,30, 35, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). Thesepolypeptides can elicit antibodies (e.g. when administered to a human)which recognise both the wild-type staphylococcal protein comprising SEQID NO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO:11. Thus the immune response will recognise both of antigens esxA andesxB. Preferred fragments of (b) provide an epitope from SEQ ID NO: 10and an epitope from SEQ ID NO: 11. The invention also provides animmunogenic composition comprising a combination of such a protein andan adjuvant, such as an aluminium hydroxide adjuvant.

The invention also provides a polypeptide comprising amino acid sequence(a) having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 241; and/or (b)comprising both a fragment of at least ‘n’ consecutive amino acids fromamino acids 1-96 of SEQ ID NO: 241 and a fragment of at least ‘n’consecutive amino acids from amino acids 103-205 of SEQ ID NO: 241,wherein ‘n’ is 7 or more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35,40, 50, 60, 70, 80, 90, 100, 150, 200, 250 or more). These polypeptides(e.g. SEQ ID NO: 250) can elicit antibodies (e.g. when administered to ahuman) which recognise both the wild-type staphylococcal proteincomprising SEQ ID NO: 10 and the wild-type staphylococcal proteincomprising SEQ ID NO: 11. Thus the immune response will recognise bothof antigens esxA and esxB. Preferred fragments of (b) provide an epitopefrom SEQ ID NO: 10 and an epitope from SEQ ID NO: 11. The invention alsoprovides an immunogenic composition comprising a combination of such aprotein and an adjuvant, such as an aluminium hydroxide adjuvant.

The invention also provides a polypeptide comprising a staphylococcalhemolysin sequence, wherein the sequence does not include a sequencehaving at least 90% identity to SEQ ID NO: 217 but can elicit antibodieswhich can kill staphylococci. The polypeptide may have a first sequencehaving 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 218 and a secondsequence having 80% or more identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to SEQ ID NO: 219, whereinthe first and second sequences are either directly joined or are joinedby an intervening amino acid sequence having fewer than 40 amino acids(e.g. ≦35 amino acids, ≦30 amino acids, ≦25 amino acids, ≦20 aminoacids, ≦15 amino acids, ≦10 amino acids, ≦5 amino acids). SEQ ID NOs:189 and 216 are examples of such polypeptides, in which the first andsecond sequences are joined by a tetrapeptide PSGS sequence (SEQ ID NO:225).

The invention also provides an immunogenic composition comprising aSta011 antigen and a Ca⁺⁺ ion. The antigen and Ca⁺⁺ ion may form acomplex e.g. atoms in the antigen may coordinate the Ca⁺⁺ ion. Theimmunogenic composition may also include an adjuvant.

The invention also provides a oligomer of a Sta011 antigen, and alsoimmunogenic compositions comprising such oligomers. The oligomer can bea dimer, trimer, tetramer, pentamer, hexamer, heptamer, octamer orhigher. An oligomer may comprise a Ca⁺⁺ ion, and a compositioncomprising Sta011 oligomers may comprise 5-500 mM Ca⁺⁺ ions.

Further Polypeptide Antigens

In additions to antigens from the various antigen groups of theinvention, immunogenic compositions may include one or more of thefollowing S. aureus antigens (or antigens comprising immunogenicfragment(s) thereof) to enhance the efficacy against S. aureus of animmune response elicited by the composition [e.g. see references 3-10]:

-   -   AhpC    -   AhpF    -   Autolysin amidase    -   Autolysin glucosaminidase    -   Collagen binding protein CAN    -   EbhB    -   GehD lipase    -   Heparin binding protein HBP (17 kDa)    -   Laminin receptor    -   MAP    -   MntC (also known as SitC)    -   MRPII    -   Npase    -   ORF0594    -   ORF0657n    -   ORF0826    -   PBP4    -   RAP (RNA III activating protein)    -   Sai-1    -   SasK    -   SBI    -   SdrG    -   SdrH    -   SSP-1    -   SSP-2    -   Vitronectin-binding protein        Combinations with Saccharides

The individual antigens identified in the antigen groups of theinvention may be used in combination with conjugated saccharideantigens. Thus the invention provides an immunogenic compositioncomprising a combination of:

-   -   (1) one or more antigen(s) selected from the first, second,        third or fourth antigen groups (as defined above); and    -   (2) one or more conjugates of a S. aureus exopolysaccharide and        a carrier protein.

A conjugate used in component (2) of this combination includes asaccharide moiety and a carrier moiety. The saccharide moiety is fromthe exopolysaccharide of S. aureus, which is a poly-N-acetylglucosamine(PNAG). The saccharide may be a polysaccharide having the size thatarises during purification of the exopolysaccharide from bacteria, or itmay be an oligosaccharide achieved by fragmentation of such apolysaccharide e.g. size can vary from over 400 kDa to between 75 and400 kDa, or between 10 and 75 kDa, or up to 30 repeat units. Thesaccharide moiety can have various degrees of N-acetylation and, asdescribed in reference 11, the PNAG may be less than 40% N-acetylated(e.g. less than 35, 30, 20, 15, 10 or 5% N-acetylated; deacetylated PNAGis also known as dPNAG). Deacetylated epitopes of PNAG can elicitantibodies that are capable of mediating opsonic killing. The PNAG mayor may not be O-succinylated e.g. it may be O-succinylated on fewer lessthan 25, 20, 15, 10, 5, 2, 1 or 0.1% of residues.

The invention also provides an immunogenic composition comprising acombination of

-   -   (1) one or more antigen(s) selected from the first, second,        third or fourth antigen groups; and    -   (2) one or more conjugates of a S. aureus capsular saccharide        and a carrier protein.

A conjugate used in component (2) of this combination includes asaccharide moiety and a carrier moiety. The saccharide moiety is fromthe capsular saccharide of a S. aureus. The saccharide may be apolysaccharide having the size that arises during purification ofcapsular polysaccharide from bacteria, or it may be an oligosaccharideachieved by fragmentation of such a polysaccharide. Capsular saccharidesmay be obtained from any suitable strain of S. aureus (or any bacteriumhaving a similar or identical saccharide), such as from a type 5 and/ora type 8 S. aureus strain and/or a type 336 S. aureus strain. Moststrains of infectious S. aureus contain either Type 5 or Type 8 capsularsaccharides. Both have FucNAcp in their repeat unit as well as ManNAcAwhich can be used to introduce a sulfhydryl group for linkage. Therepeating unit of the Type 5 saccharide is →4)-β-D-ManNAcA-(1→4)-α-L-FucNAc(3OAc)-(1→3)-β-D-FucNAc-(1→, whereas the repeatingunit of the Type 8 saccharide is→3)-β-D-ManNAcA(4OAc)-(1→3)-α-L-FucNAc(1→3)-α-D-FucNAc(1→. The type 336saccharide is a O-linked hexosamine with no O-acetylation [12,13] and iscross-reactive with antibodies raised against the 336 strain (ATCC55804). A combination of a type 5 and a type 8 saccharide is typical,and a type 336 saccharide may be added to this pairing [14].

The invention also provides an immunogenic composition comprising acombination of

-   -   (1) one or more antigen(s) selected from the first, second,        third or fourth antigen groups;    -   (2) one or more conjugates of a S. aureus exopolysaccharide and        a carrier protein; and    -   (3) one or more conjugates of a S. aureus capsular saccharide        and a carrier protein.

The carrier moiety in these conjugates will usually be a protein, butusually not one of the antigens of (1). Typical carrier proteins arebacterial toxins, such as diphtheria or tetanus toxins, or toxoids ormutants or fragments thereof. The CRM197 diphtheria toxin mutant [15] isuseful. Other suitable carrier proteins include the N. meningitidisouter membrane protein complex [16], synthetic peptides [17,18], heatshock proteins [19,20], pertussis proteins [21,22], cytokines [23],lymphokines [23], hormones [23], growth factors [23], artificialproteins comprising multiple human CD4⁺ T cell epitopes from variouspathogen-derived antigens [24] such as N19 [25], protein D from H.influenzae [26-28], pneumolysin [29] or its non-toxic derivatives [30],pneumococcal surface protein PspA [31], iron-uptake proteins [32], toxinA or B from C. difficile [33], recombinant P. aeruginosa exoprotein A(rEPA) [34], etc. In some embodiments the carrier protein is a S. aureusprotein, such as an antigen selected from the first, second, third orfourth antigen groups.

Where a composition includes more than one conjugate, each conjugate mayuse the same carrier protein or a different carrier protein.

Conjugates may have excess carrier (w/w) or excess saccharide (w/w). Insome embodiments, a conjugate may include substantially equal weights ofeach.

The carrier molecule may be covalently conjugated to the carrierdirectly or via a linker. Direct linkages to the protein may be achievedby, for instance, reductive amination between the saccharide and thecarrier, as described in, for example, references 35 and 36. Thesaccharide may first need to be activated e.g. by oxidation. Linkagesvia a linker group may be made using any known procedure, for example,the procedures described in references 37 and 38. A preferred type oflinkage is an adipic acid linker, which may be formed by coupling a free—NH₂ group (e.g. introduced to a glucan by amination) with adipic acid(using, for example, diimide activation), and then coupling a protein tothe resulting saccharide-adipic acid intermediate [39,40]. Anotherpreferred type of linkage is a carbonyl linker, which may be formed byreaction of a free hydroxyl group of a saccharide CDI [41, 42] followedby reaction with a protein to form a carbamate linkage. Other linkersinclude β-propionamido [43], nitrophenyl-ethylamine [44], haloacylhalides [45], glycosidic linkages [46], 6-aminocaproic acid [47], ADH[48], C₄ to C₁₂ moieties [49], etc. Carbodiimide condensation can alsobe used [50].

PNAG conjugates may be prepared in various ways e.g. by a processcomprising: a) activating the PNAG by adding a linker comprising amaleimide group to form an activated PNAG; b) activating the carrierprotein by adding a linker comprising a sulphydryl group to form anactivated carrier protein; and c) reacting the activated PNAG and theactivated carrier protein to form a PNAG-carrier protein conjugate; orby a process comprising a) activating the PNAG by adding a linkercomprising a sulphydryl group to form an activated PNAG; b) activatingthe carrier protein by adding a linker comprising a maleimide group toform an activated carrier protein; and c) reacting the activated PNAGand the activated carrier protein to form a PNAG-carrier proteinconjugate; or by a process comprising a) activating the PNAG by adding alinker comprising a sulphydryl group to form an activated PNAG; b)activating the carrier protein by adding a linker comprising asulphydryl group to form an activated carrier protein; and c) reactingthe activated PNAG and the activated carrier protein to form aPNAG-carrier protein conjugate.

The individual antigens identified in the antigen groups of theinvention may be used as carrier proteins for exopolysaccharides, toform a covalent conjugate. Thus the invention provides an immunogeniccomposition comprising a conjugate of (1) an antigen selected from thefirst, second, third and fourth antigen groups and (2) a S. aureusexopolysaccharide. The invention also provides an immunogeniccomposition comprising a conjugate of (1) an antigen selected from thefirst, second, third and fourth antigen groups and (2) a S. aureuscapsular saccharide. Further characteristics of such conjugates aredescribed above. These conjugates may be combined with any of theantigens disclosed herein.

Combinations with Non-Staphylococcal Antigens

The individual antigens identified in the antigen groups of theinvention may be used in combination with non-staphylococcal antigens,and in particular with antigens from bacteria associated with nosocomialinfections. Thus the invention provides an immunogenic compositioncomprising a combination of:

-   -   (1) one or more antigen(s) selected from the first, second,        third and fourth antigen groups (as defined above); and    -   (2) one or more antigen(s) selected from the group consisting        of: Clostridium difficile; Pseudomonas aeruginosa; Candida        albicans; and extraintestinal pathogenic Escherichia coli.

Further suitable antigens for use in combination with staphylococcalantigens of the invention are listed on pages 33-46 of reference 51.

First Antigen Group

clfA

The ‘clfA’ antigen is annotated as ‘clumping factor A’. In the NCTC 8325strain clfA is SAOUHSC_(—)00812 and has amino acid sequence SEQ ID NO: 1(GI:88194572). In the Newman strain it is nwmn_(—)0756 (GI:151220968).

Useful clfA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 1 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 1; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 1, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These clfA proteins include variants of SEQ ID NO: 1.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 1. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 1 while retaining at least one epitopeof SEQ ID NO: 1. The final 368 C-terminal amino acids of SEQ ID NO: 1can usefully be omitted. The first 39 N-terminal amino acids of SEQ IDNO: 1 can usefully be omitted. Other fragments omit one or more proteindomains.

SEQ ID NO: 224 is a useful fragment of SEQ ID NO: 1 (‘ClfA₄₀₋₅₅₉’). Thisfragments omits the long repetitive region towards the C-terminal of SEQID NO: 1.clfB

The ‘clfB’ antigen is annotated as ‘clumping factor B’. In the NCTC 8325strain clfB is SAOUHSC_(—)02963 and has amino acid sequence SEQ ID NO: 2(GI:88196585). In the Newman strain it is nwmn_(—)2529 (GI:151222741).

Useful clfB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 2 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 2; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 2, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These clfB proteins include variants of SEQ ID NO: 2.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 2. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 2 while retaining at least one epitopeof SEQ ID NO: 2. The final 40 C-terminal amino acids of SEQ ID NO: 2 canusefully be omitted. The first 44 N-terminal amino acids of SEQ ID NO: 2can usefully be omitted. Other fragments omit one or more proteindomains. ClfB is naturally a long protein and so the use of fragments ishelpful e.g. for purification, handling, fusion, expression, etc. SEQ IDNO: 163 is a useful fragment of SEQ ID NO: 2 (‘ClfB₄₅₋₅₅₂’). Thisfragment includes the most exposed domain of ClfB and is more easilyused at an industrial scale. It also reduces the antigen's similaritywith human proteins. Other useful fragments, based on a 3-domain modelof ClfB, include: ClfB₄₅₋₃₆₀ (also known as ClfB-N12; SEQ ID NO: 196);ClfB₂₁₂₋₅₄₂ (also known as CLfB-N23; SEQ ID NO: 197); and ClfB₃₆₀₋₅₄₂(also known as CLfB-N3; SEQ ID NO: 198).

coA

The ‘coA’ antigen is annotated as ‘coagulase Coa’. In the NCTC 8325strain coA is SAOUHSC_(—)00192 and has amino acid sequence SEQ ID NO: 3(GI:88194002). In the Newman strain it is nwmn_(—)0166 (GI:151220378).

Useful coA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 3 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 3; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 3, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These coA proteins include variants of SEQ ID NO: 3.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 3. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 3 while retaining at least one epitopeof SEQ ID NO: 3. The first 14 N-terminal amino acids of SEQ ID NO: 3 canusefully be omitted. Other fragments omit one or more protein domains.

eap

The ‘eap’ antigen is annotated as ‘MHC class II analog protein’. In theNCTC 8325 strain eap is SAOUHSC_(—)02161 and has amino acid sequence SEQID NO: 4 (GI:88195840). In the Newman strain it is nwmn_(—)1872(GI:151222084).

Useful eap antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 4 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 4; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 4, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These eap proteins include variants of SEQ ID NO: 4.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 4. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 4 while retaining at least one epitopeof SEQ ID NO: 4. The first 17 N-terminal amino acids of SEQ ID NO: 4 canusefully be omitted. Other fragments omit one or more protein domains.

ebhA

The ‘ebhA’ antigen is annotated as ‘EbhA’. In the NCTC 8325 strain ebhAis SAOUHSC_(—)01447 and has amino acid sequence SEQ ID NO: 5(GI:88195168).

Useful ebhA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 5 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 5; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 5, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These ebhA proteins include variants of SEQ ID NO: 5.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 5. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 5 while retaining at least one epitopeof SEQ ID NO: 5. The first 39 N-terminal amino acids of SEQ ID NO: 5 canusefully be omitted. Other fragments omit one or more protein domains.

ebpS

The ‘ebpS’ antigen is annotated as ‘elastin binding protein EbpS’. Inthe NCTC 8325 strain ebpS is SAOUHSC_(—)01501 and has amino acidsequence SEQ ID NO: 6 (GI:88195217). In the Newman strain it isnwmn_(—)1389 (GI:151221601).

Useful ebpS antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 6 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 6; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 6, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These ebpS proteins include variants of SEQ ID NO: 6.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 6. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 6 while retaining at least one epitopeof SEQ ID NO: 6. Other fragments omit one or more protein domains.

SEQ ID NO: 165 is a useful fragment of SEQ ID NO: 6 (‘EbpS₁₋₁₉₈’). Thisfragment includes the most exposed domain of EbpS and is more easilyused at an industrial scale. It also reduces the antigen's similaritywith human proteins.

efb

The ‘efb’ antigen is annotated as ‘fibrinogen-binding proteintruncated’. In the NCTC 8325 strain efb is SAOUHSC_(—)01114 and hasamino acid sequence SEQ ID NO: 7 (GI:88194860). In the Newman strain itis nwmn_(—)1069 (GI:151221281).

Useful efb antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 7 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 7; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 7, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These efb proteins include variants of SEQ ID NO: 7. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 7. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 7 while retaining at least one epitope of SEQID NO: 7. The first 14 N-terminal amino acids of SEQ ID NO: 7 canusefully be omitted. Other fragments omit one or more protein domains.

emp

The ‘emp’ antigen is annotated as ‘extracellular matrix and plasmabinding protein’. In the NCTC 8325 strain emp is SAOUHSC_(—)00816 andhas amino acid sequence SEQ ID NO: 8 (GI:88194575). In the Newman strainit is nwmn_(—)0758 (GI:151220970).

Useful emp antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 8 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 8; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 8, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These emp proteins include variants of SEQ ID NO: 8.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 8. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 8 while retaining at least one epitopeof SEQ ID NO: 8. The first 26 N-terminal amino acids of SEQ ID NO: 8 canusefully be omitted. Other fragments omit one or more protein domains.

SEQ ID NOs: 190, 191, 192 and 193 are useful fragments of SEQ ID NO: 8(‘Emp₃₅₋₃₄₀’, ‘Emp₂₇₋₃₃₄’, ‘Emp₃₅₋₃₃₄’ and ‘Emp₂₇₋₁₄₇’, respectively).

esaC

The ‘esaC’ antigen is annotated as ‘esaC’. In the NCTC 8325 strain esaCis SAOUHSC_(—)00264 and has amino acid sequence SEQ ID NO: 9(GI:88194069).

Useful esaC antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 9 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 9; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 9, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 ormore). These esaC proteins include variants of SEQ ID NO: 9. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 9. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 9 while retaining at least one epitope of SEQID NO: 9. Other fragments omit one or more protein domains.

esxA

The ‘esxA’ antigen is annotated as ‘protein’. In the NCTC 8325 strainesxA is SAOUHSC_(—)00257 and has amino acid sequence SEQ ID NO: 10(GI:88194063).

Useful esxA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 10 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 10; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 10, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90 or more).These esxA proteins include variants of SEQ ID NO: 10. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 10. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 10 while retaining at least one epitope of SEQID NO: 10. Other fragments omit one or more protein domains.

esxB

The ‘esxB’ antigen is annotated as ‘esxB’. In the NCTC 8325 strain esxBis SAOUHSC_(—)00265 and has amino acid sequence SEQ ID NO: 11(GI:88194070).

Useful esxB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 11 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 11; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 11, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 ormore). These esxB proteins include variants of SEQ ID NO: 11. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 11. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 11 while retaining at least one epitope of SEQID NO: 11. Other fragments omit one or more protein domains.

FnBA

The ‘FnBA’ antigen is annotated as ‘fibronectin-binding protein Aprecursor FnBPA’. In the NCTC 8325 strain FnBA is SAOUHSC_(—)02803 andhas amino acid sequence SEQ ID NO: 12 (GI:88196438). In the Newmanstrain it is nwmn_(—)2399 (GI:151222611). Proteomic analysis hasrevealed that this protein is secreted or surface-exposed.

Useful FnBA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 12 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 12; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 12, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These FnBA proteins include variants of SEQ ID NO:12. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 12.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 12 while retaining at least oneepitope of SEQ ID NO: 12. The final 37 C-terminal amino acids of SEQ IDNO: 12 can usefully be omitted. Other fragments omit one or more proteindomains. FnBA is naturally a long protein and so the use of fragments ishelpful e.g. for purification, handling, fusion, expression, etc.

SEQ ID NOs: 166 (‘FnBA₁₋₅₁₁’) and 167 (‘FnBA₅₁₂₋₉₅₃’) are usefulfragments of SEQ ID NO: 12. These fragments are more easily used at anindustrial scale.

FnBB

The ‘FnBB’ antigen is annotated as ‘fibronectin binding protein BFnBPB’. In the NCTC 8325 strain FnBB is SAOUHSC_(—)02802 and has aminoacid sequence SEQ ID NO: 13 (GI:88196437). In the Newman strain it isnwmn_(—)2397 (GI:151222609).

Useful FnBB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 13 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 13; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 13, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These FnBB proteins include variants of SEQ ID NO:13. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 13.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 13 while retaining at least oneepitope of SEQ ID NO: 13. The final 37 C-terminal amino acids of SEQ IDNO: 13 can usefully be omitted. Other fragments omit one or more proteindomains.

Hla

The ‘Hla’ antigen is the ‘alpha-hemolysin precursor’ also known as‘alpha toxin’ or simply ‘hemolysin’. In the NCTC 8325 strain Hla isSAOUHSC_(—)01121 and has amino acid sequence SEQ ID NO: 14(GI:88194865). In the Newman strain it is nwmn_(—)1073 (GI:151221285).Hla is an important virulence determinant produced by most strains of S.aureus, having pore-forming and haemolytic activity. Anti-Hla antibodiescan neutralise the detrimental effects of the toxin in animal models,and Hla is particularly useful for protecting against pneumonia.

Useful Hla antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 14 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 14; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 14, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These Hla proteins include variants of SEQ ID NO: 14.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 14. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 14 while retaining at least oneepitope of SEQ ID NO: 14. The first 26 N-terminal amino acids of SEQ IDNO: 14 can usefully be omitted (e.g. to give SEQ ID NO: 231). Truncationat the C-terminus can also be used e.g. leaving only 50 amino acids(residues 27-76 of SEQ ID NO: 14) [52]. Other fragments omit one or moreprotein domains.

Hla's toxicity can be avoided in compositions of the invention bychemical inactivation (e.g. using formaldehyde, glutaraldehyde or othercross-linking reagents). Instead, however, it is preferred to use mutantforms of Hla which remove its toxic activity while retaining itsimmunogenicity. Such detoxified mutants are already known in the art.One useful Hla antigen has a mutation at residue 61 of SEQ ID NO: 14,which is residue 35 of the mature antigen (i.e. after omitting the first26 N-terminal amino acids=residue 35 of SEQ ID NO: 231). Thus residue 61may not be histidine, and may instead be e.g. Ile, Val or preferablyLeu. A His-Arg mutation at this position can also be used. For example,SEQ ID NO: 150 is the mature mutant Hla-H35L sequence (i.e. SEQ ID NO:231 with a H35L mutation) and a useful Hla antigen comprises SEQ ID NO:150. Another useful mutation replaces a long loop with a short sequencee.g. to replace the 39mer at residues 136-174 of SEQ ID NO: 14 with atetramer such as PSGS (SEQ ID NO: 225), as in SEQ ID NO: 189 (which alsoincludes the H35L mutation) and SEQ ID NO: 216 (which does not includethe H35L mutation). Another useful mutation replaces residue Y101 e.g.with a leucine (SEQ ID NO: 242). Another useful mutation replacesresidue D152 e.g. with a leucine (SEQ ID NO: 243). Another useful mutantreplaces residues H35 and Y101 e.g. with a leucine (SEQ ID NO: 244).Another useful mutant replaces residues H35 and D152 e.g. with a leucine(SEQ ID NO: 245).

Further useful Hla antigens are disclosed in references 53 and 54.

SEQ ID NOs: 160, 161 & 194 are three useful fragments of SEQ ID NO: 14(‘Hla₂₇₋₇₆’, ‘Hla₂₇₋₈₉’ and ‘Hla₂₇₋₇₉’, respectively). SEQ ID NOs: 158,159 and 195 are the corresponding fragments from SEQ ID NO: 150.

One useful Hla sequence is SEQ ID NO: 232, which was used in theexamples. It has a N-terminal Met, then an Ala-Ser dipeptide from theexpression vector, then SEQ ID NO: 150 (from NCTC8325 strain). It isencoded by SEQ ID NO: 233.

hlgB

The ‘hlgB’ antigen is annotated as ‘leukocidin f subunit precursorHlgB’. In the NCTC 8325 strain hlgB is SAOUHSC_(—)02710 and has aminoacid sequence SEQ ID NO: 15 (GI:88196350).

Useful hlgB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 15 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 15; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 15, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These hlgB proteins include variants of SEQ ID NO:15. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 15.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 15 while retaining at least oneepitope of SEQ ID NO: 15. The first 26 N-terminal amino acids of SEQ IDNO: 15 can usefully be omitted. Other fragments omit one or more proteindomains.

hlgC

The ‘hlgC’ antigen is annotated as ‘leukocidin s subunit precursorHlgC’. In the NCTC 8325 strain hlgC is SAOUHSC_(—)02709 and has aminoacid sequence SEQ ID NO: 16 (GI:88196349).

Useful hlgC antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 16 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 16; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 16, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These hlgC proteins include variants of SEQ ID NO:16. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 16.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 16 while retaining at least oneepitope of SEQ ID NO: 16. The first 29 N-terminal amino acids of SEQ IDNO: 16 can usefully be omitted. Other fragments omit one or more proteindomains.

isdA

The ‘isdA’ antigen is annotated as ‘IsdA protein’. In the NCTC 8325strain isdA is SAOUHSC_(—)01081 and has amino acid sequence SEQ ID NO:17 (GI:88194829). In the Newman strain it is nwmn_(—)1041(GI:151221253).

Useful isdA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 17 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 17; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 17, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These isdA proteins include variants of SEQ ID NO:17. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 17.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 17 while retaining at least oneepitope of SEQ ID NO: 17. The final 38 C-terminal amino acids of SEQ IDNO: 17 can usefully be omitted. The first 46 N-terminal amino acids ofSEQ ID NO: 17 can usefully be omitted. Truncation to exclude theC-terminal 38mer of SEQ ID NO: 17 (beginning with the LPKTG motif) isalso useful. Other fragments omit one or more protein domains.

SEQ ID NO: 157 is a useful fragment of SEQ ID NO: 17 (amino acids 40-184of SEQ ID NO: 17; ‘IsdA₄₀₋₁₈₄’) which includes the natural protein'sheme binding site and includes the antigen's most exposed domain. Italso reduces the antigen's similarity with human proteins. Other usefulfragments are disclosed in references 55 and 56.

IsdA does not adsorb well to aluminium hydroxide adjuvants, so IsdApresent in a composition may me unadsorbed or may be adsorbed to analternative adjuvant e.g. to an aluminium phosphate.

Anti-IsdA antibodies protect mice against S. aureus abscess formationand lethal challenge [57].

isdB

The ‘isdB’ antigen is annotated as ‘neurofilament protein isdB’. In theNCTC 8325 strain isdB is SAOUHSC_(—)01079 and has amino acid sequenceSEQ ID NO: 18 (GI:88194828). IsdB has been proposed for use as a vaccineantigen on its own [2], but this may not prevent pneumonia.

Useful isdB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 18 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 18; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 18, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These isdB proteins include variants of SEQ ID NO:18. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 18.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 18 while retaining at least oneepitope of SEQ ID NO: 18. The final 36 C-terminal amino acids of SEQ IDNO: 18 can usefully be omitted. The first 40 N-terminal amino acids ofSEQ ID NO: 18 can usefully be omitted. Other fragments omit one or moreprotein domains. Useful fragments of IsdB are disclosed in references 56and 58 e.g. lacking 37 internal amino acids of SEQ ID 18.

Anti-IsdB antibodies protect mice against S. aureus abscess formationand lethal challenge [57].

In some embodiments, compositions of the invention do not include anisdB antigen.

isdC

The ‘isdC’ antigen is annotated as ‘protein’. In the NCTC 8325 strainisdC is SAOUHSC_(—)01082 and has amino acid sequence SEQ ID NO: 19(GI:88194830).

Useful isdC antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 19 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 19; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 19, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These isdC proteins include variants of SEQ ID NO: 19.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 19. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 19 while retaining at least oneepitope of SEQ ID NO: 19. The final 39 C-terminal amino acids of SEQ IDNO: 19 can usefully be omitted. The first 28 N-terminal amino acids ofSEQ ID NO: 19 can usefully be omitted. Other fragments omit one or moreprotein domains. Useful fragments of IsdB are disclosed in reference 56.

Reference 59 discloses antigens which usefully include epitopes fromboth IsdB and IsdH.

isdG

The ‘isdG’ antigen is annotated as ‘heme-degrading monooxygenase IsdG’.In the NCTC 8325 strain isdG is SAOUHSC_(—)01089 and has amino acidsequence SEQ ID NO: 20 (GI:88194836).

Useful isdG antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 20 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 20; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 20, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 ormore). These isdG proteins include variants of SEQ ID NO: 20. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 20. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 20 while retaining at least one epitope of SEQID NO: 20. Other fragments omit one or more protein domains.

isdH

The ‘isdH’ antigen is annotated as ‘isdH’. In the NCTC 8325 strain isdHis SAOUHSC_(—)01843 and has amino acid sequence SEQ ID NO: 21(GI:88195542). In the Newman strain it is nwmn_(—)1624 (GI:151221836).It has also been known as HarA.

Useful isdH antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 21 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 21; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 21, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These isdH proteins include variants of SEQ ID NO:21. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 21.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 21 while retaining at least oneepitope of SEQ ID NO: 21. The final 35 C-terminal amino acids of SEQ IDNO: 21 can usefully be omitted. The first 40 N-terminal amino acids ofSEQ ID NO: 21 can usefully be omitted. Other fragments omit one or moreprotein domains.

Reference 59 discloses antigens which usefully include epitopes fromboth IsdB and IsdH.

isdI

The ‘isdI’ antigen is annotated as ‘heme-degrading monooxygenase IsdI’.In the NCTC 8325 strain isdI is SAOUHSC_(—)00130 and has amino acidsequence SEQ ID NO: 22 (GI:88193943).

Useful isdI antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 22 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 22; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 22, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 ormore). These isdI proteins include variants of SEQ ID NO: 22. Preferredfragments of (b) comprise an epitope from SEQ ID NO: 22. Other preferredfragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 15, 20, 25 or more) from the C-terminus and/or one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theN-terminus of SEQ ID NO: 22 while retaining at least one epitope of SEQID NO: 22. Other fragments omit one or more protein domains.

lukD

The ‘lukD’ antigen is annotated as ‘leukotoxin LukD’. In the NCTC 8325strain lukD is SAOUHSC_(—)01954 and has amino acid sequence SEQ ID NO:23 (GI:88195647). In the Newman strain it is nwmn_(—)1718(GI:151221930).

Useful lukD antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 23 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 23; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 23, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These lukD proteins include variants of SEQ ID NO:23. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 23.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 23 while retaining at least oneepitope of SEQ ID NO: 23. The final 43 C-terminal amino acids of SEQ IDNO: 23 can usefully be omitted. The first 26 N-terminal amino acids ofSEQ ID NO: 23 can usefully be omitted. Other fragments omit one or moreprotein domains.

lukE

The ‘lukE’ antigen is annotated as ‘leukotoxin LukE’. In the NCTC 8325strain lukE is SAOUHSC_(—)01955 and has amino acid sequence SEQ ID NO:24 (GI:88195648).

Useful lukE antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 24 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 24; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 24, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These lukE proteins include variants of SEQ ID NO:24. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 24.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 24 while retaining at least oneepitope of SEQ ID NO: 24. Other fragments omit one or more proteindomains.

lukF

The ‘lukF’ antigen is annotated as ‘Leukocidin/Hemolysin toxin familyLukF’. In the NCTC 8325 strain lukF is SAOUHSC_(—)02241 and has aminoacid sequence SEQ ID NO: 25 (GI:88195914).

Useful lukF antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 25 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 25; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 25, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These lukF proteins include variants of SEQ ID NO:25. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 25.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 25 while retaining at least oneepitope of SEQ ID NO: 25. Other fragments omit one or more proteindomains.

lukS

The ‘lukS’ antigen is annotated as ‘probable leukocidin S subunit LukS’.In the NCTC 8325 strain lukS is SAOUHSC_(—)02243 and has amino acidsequence SEQ ID NO: 26 (GI:88195915). In the Newman strain it isnwmn_(—)1928 (GI:151222140).

Useful lukS antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 26 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 26; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 26, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These lukS proteins include variants of SEQ ID NO:26. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 26.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 26 while retaining at least oneepitope of SEQ ID NO: 26. The first 22 N-terminal amino acids of SEQ IDNO: 26 can usefully be omitted. Other fragments omit one or more proteindomains.

nuc

The ‘nuc’ antigen is annotated as ‘thermonuclease precursor’. In theNCTC 8325 strain nuc is SAOUHSC_(—)01316 and has amino acid sequence SEQID NO: 27 (GI:88195046).

Useful nuc antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 27 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 27; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 27, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These nuc proteins include variants of SEQ ID NO: 27.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 27. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 27 while retaining at least oneepitope of SEQ ID NO: 27. The final 39 C-terminal amino acids of SEQ IDNO: 27 can usefully be omitted. The first 19 N-terminal amino acids ofSEQ ID NO: 27 can usefully be omitted. Other fragments omit one or moreprotein domains.

sasA

The ‘sasA’ antigen is annotated as ‘SasA’. In the NCTC 8325 strain sasAis SAOUHSC_(—)02990 and has amino acid sequence SEQ ID NO: 28(GI:88196609).

Useful sasA antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 28 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 28; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 28, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sasA proteins include variants of SEQ ID NO:28. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 28.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 28 while retaining at least oneepitope of SEQ ID NO: 28. The final 43 C-terminal amino acids of SEQ IDNO: 28 can usefully be omitted. The first 90 N-terminal amino acids ofSEQ ID NO: 28 can usefully be omitted. Other fragments omit one or moreprotein domains.

sasB

The ‘sasB’ antigen is annotated as ‘fmtB protein; SasB’. In the NCTC8325 strain sasB is SAOUHSC_(—)02404 and has amino acid sequence SEQ IDNO: 29 (GI:88196065).

Useful sasB antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 29 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 29; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 29, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sasB proteins include variants of SEQ ID NO:29. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 29.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 29 while retaining at least oneepitope of SEQ ID NO: 29. The final 39 C-terminal amino acids of SEQ IDNO: 29 can usefully be omitted. The first 38 N-terminal amino acids ofSEQ ID NO: 29 can usefully be omitted. Other fragments omit one or moreprotein domains.

sasC

The ‘sasC’ antigen is annotated as ‘Mrp protein; SasC’. In the NCTC 8325strain sasC is SAOUHSC_(—)01873 and has amino acid sequence SEQ ID NO:30 (GI:88195570).

Useful sasC antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 30 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 30; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 30, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sasC proteins include variants of SEQ ID NO:30. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 30.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 30 while retaining at least oneepitope of SEQ ID NO: 30. The final 36 C-terminal amino acids of SEQ IDNO: 30 can usefully be omitted. The first 37 N-terminal amino acids ofSEQ ID NO: 30 can usefully be omitted. Other fragments omit one or moreprotein domains.

sasD

The ‘sasD’ antigen is annotated as ‘SasD protein’. In the NCTC 8325strain sasD is SAOUHSC_(—)00094 and has amino acid sequence SEQ ID NO:31 (GI:88193909).

Useful sasD antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 31 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 31; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 31, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These sasD proteins include variants of SEQ ID NO: 31.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 31. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 31 while retaining at least oneepitope of SEQ ID NO: 31. The first 28 N-terminal amino acids of SEQ IDNO: 31 can usefully be omitted. Other fragments omit one or more proteindomains.

sasF

The ‘sasF’ antigen is annotated as ‘sasF protein’. In the NCTC 8325strain sasF is SAOUHSC_(—)02982 and has amino acid sequence SEQ ID NO:32 (GI:88196601).

Useful sasF antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 32 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 32; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 32, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sasF proteins include variants of SEQ ID NO:32. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 32.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 32 while retaining at least oneepitope of SEQ ID NO: 32. The final 39 C-terminal amino acids of SEQ IDNO: 32 can usefully be omitted. The first 37 N-terminal amino acids ofSEQ ID NO: 32 can usefully be omitted. Other fragments omit one or moreprotein domains.

sdrC

The ‘sdrC’ antigen is annotated as ‘sdrC protein’. In the NCTC 8325strain sdrC is SAOUHSC_(—)00544 and has amino acid sequence SEQ ID NO:33 (GI:88194324).

Useful sdrC antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 33 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 33; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 33, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sdrC proteins include variants of SEQ ID NO:33. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 33.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 33 while retaining at least oneepitope of SEQ ID NO: 33. The final 38 C-terminal amino acids of SEQ IDNO: 33 can usefully be omitted. The first 50 N-terminal amino acids ofSEQ ID NO: 33 can usefully be omitted. Other fragments omit one or moreprotein domains. SdrC is naturally a long protein and so the use offragments is helpful e.g. for purification, handling, fusion,expression, etc.

SEQ ID NO: 164 is a useful fragment of SEQ ID NO: 33 (‘SdrC5₁₋₅₁₈’).This fragment includes the most exposed domain of SdrC and is moreeasily used at an industrial scale. It also reduces the antigen'ssimilarity with human proteins.

sdrD

The ‘sdrD’ antigen is annotated as ‘sdrD protein’. In the NCTC 8325strain sdrD is SAOUHSC_(—)00545 and has amino acid sequence SEQ ID NO:34 (GI:88194325).

Useful sdrD antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 34 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 34; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 34, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sdrD proteins include variants of SEQ ID NO:34. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 34.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 34 while retaining at least oneepitope of SEQ ID NO: 34. The final 38 C-terminal amino acids of SEQ IDNO: 34 can usefully be omitted. The first 52 N-terminal amino acids ofSEQ ID NO: 34 can usefully be omitted. Other fragments omit one or moreprotein domains. SdrD is naturally a long protein and so the use offragments is very helpful e.g. for purification, handling, fusion,expression, etc.

SEQ ID NO: 156 is a useful fragment of SEQ ID NO: 34 (‘SdrD₅₃₋₅₉₂’).This fragment includes the most exposed domain of SdrD and is moreeasily used at an industrial scale. It also reduces the antigen'ssimilarity with human proteins. Another useful fragment, with the sameC-terminus residue, is SdrD₃₉₄₋₅₉₂ (also known as SdrD-N3; SEQ ID NO:199). Another useful fragment is SEQ ID NO: 236 (amino acids 593-1123 ofSEQ ID NO: 34), referred to herein as ‘SdrD_(cnaB)’.

sdrE2

The ‘sdrE2’ antigen is annotated as ‘Ser-Asp rich fibrinogen/bonesialoprotein-binding protein SdrE’. In the Newman strain sdrE2 isNWMN_(—)0525 and has amino acid sequence SEQ ID NO: 35 (GI:151220737).

Useful sdrE2 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 35 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 35; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 35, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sdrE2 proteins include variants of SEQ ID NO:35. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 35.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 35 while retaining at least oneepitope of SEQ ID NO: 35. The final 38 C-terminal amino acids of SEQ IDNO: 35 can usefully be omitted. The first 52 N-terminal amino acids ofSEQ ID NO: 35 can usefully be omitted. Other fragments omit one or moreprotein domains. SdrE2 is naturally a long protein and so the use offragments is very helpful e.g. for purification, handling, fusion,expression, etc.

SEQ ID NO: 155 is a useful fragment of SEQ ID NO: 35 (‘SdrE₅₃₋₆₃₂’).This fragment includes the most exposed domain of SdrE2 and is moreeasily used at an industrial scale. It also reduces the antigen'ssimilarity with human proteins.

spa

The ‘spa’ antigen is annotated as ‘protein A’ or ‘SpA’. In the NCTC 8325strain spa is SAOUHSC_(—)00069 and has amino acid sequence SEQ ID NO: 36(GI:88193885). In the Newman strain it is nwmn_(—)0055 (GI:151220267).All S. aureus strains express the structural gene for spa, a wellcharacterized virulence factor whose cell wall-anchored surface proteinproduct has five highly homologous immunoglobulin binding domainsdesignated E, D, A, B, and C [60]. These domains display ˜80% identityat the amino acid level, are 56 to 61 residues in length, and areorganized as tandem repeats [61]. SpA is synthesized as a precursorprotein with an N-terminal signal peptide and a C-terminal sortingsignal [62,63]. Cell wall-anchored spa is displayed in great abundanceon the staphylococcal surface [64,65]. Each of its immunoglobulinbinding domains is composed of anti-parallel α-helices that assembleinto a three helix bundle and can bind the Fc domain of immunoglobulin G(IgG) [66,67], the VH3 heavy chain (Fab) of IgM (i.e. the B cellreceptor) [68], the von Willebrand factor at its A1 domain [69] and/orthe TNF-α receptor I (TNFRI) [70], which is displayed on surfaces ofairway epithelia.

Useful spa antigens can elicit an antibody (e.g. when administered to ahuman) that recognises SEQ ID NO: 36 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 36; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 36, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These spa proteins include variants of SEQ ID NO: 36.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 36. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 36 while retaining at least oneepitope of SEQ ID NO: 36. The final 35 C-terminal amino acids of SEQ IDNO: 36 can usefully be omitted. The first 36 N-terminal amino acids ofSEQ ID NO: 36 can usefully be omitted. Other fragments omit one or moreprotein domains. Reference 71 suggests that individual IgG-bindingdomains might be useful immunogens, alone or in combination.

SEQ ID NO: 162 is a useful fragment of SEQ ID NO: 36 (‘Spa₃₇₋₃₂₅’). Thisfragment contains all the five SpA Ig-binding domains (which arenaturally arranged from N- to C-terminus in the order E, D, A, B, C) andincludes the most exposed domain of SpA. It also reduces the antigen'ssimilarity with human proteins. Other useful fragments may omit 1, 2, 3or 4 of the natural A, B, C, D and/or E domains to prevent the excessiveB cell expansion and then apoptosis which might occur if spa functionsas a B cell superantigen. As reported in reference 71, other usefulfragments may include only 1, 2, 3 or 4 of the natural A, B, C, D and/orE domains e.g. comprise only the SpA(A) domain but not B to E, orcomprise only the SpA(D) domain but not A, B, C or E, etc. Thus a spaantigen useful with the invention may include 1, 2, 3, 4 or 5IgG-binding domains, but ideally has 4 or fewer

If an antigen includes only one type of spa domain (e.g. only the Spa(A)or SpA(D) domain), it may include more than one copy of this domain e.g.multiple SpA(D) domains in a single polypeptide chain.

An individual domain within the antigen may be mutated at 1, 2, 3, 4, 5,6, 7, 8, 9, 10 or more amino acids relative to SEQ ID NO: 36 (e.g. seeref. 71, disclosing mutations at residues 3 and/or 24 of domain D, atresidue 46 and/or 53 of domain A, etc.). Such mutants should not removethe antigen's ability to elicit an antibody that recognises SEQ ID NO:36, but may remove the antigen's binding to IgG and/or other humanproteins (such as human blood proteins).

In certain aspects a spa antigen includes a substitution at (a) one ormore amino acid substitution in an IgG Fc binding sub-domain of SpAdomain A, B, C, D and/or E that disrupts or decreases binding to IgG Fc,and (b) one or more amino acid substitution in a V_(H)3 bindingsub-domain of SpA domain A, B, C, D, and/or E that disrupts or decreasesbinding to V_(H)3. In certain embodiments, a variant SpA comprises atleast or at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more variant SpAdomain D peptides.

Second Antigen Group

sta001

The ‘sta001’ antigen is annotated as ‘5’-nucleotidase family protein'.In the NCTC 8325 strain sta001 is SAOUHSC_(—)00025 and has amino acidsequence SEQ ID NO: 37 (GI:88193846). In the Newman strain it isnwmn_(—)0022 (GI:151220234). It has also been referred to as AdsA andSasH and SA0024.

Useful sta001 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 37 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 37; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 37, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta001 proteins include variants of SEQ ID NO:37. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 37.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 37 while retaining at least oneepitope of SEQ ID NO: 37. The final 34 C-terminal amino acids of SEQ IDNO: 37 can usefully be omitted. The first 38 N-terminal amino acids ofSEQ ID NO: 37 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta002

The ‘sta002’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta002 is SAOUHSC_(—)00356 and has amino acid sequence SEQ ID NO:38 (GI:88194155). In the Newman strain it is nwmn_(—)0364(GI:151220576).

Useful sta002 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 38 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 38; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 38, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These sta002 proteins include variants of SEQ ID NO: 38.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 38. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 38 while retaining at least oneepitope of SEQ ID NO: 38. The first 18 N-terminal amino acids of SEQ IDNO: 38 can usefully be omitted. Other fragments omit one or more proteindomains.

SEQ ID NOs: 153 (‘Sta002₁₉₋₁₈₇’) and 154 (‘sta002₁₉₋₁₂₄’) are two usefulfragments of SEQ ID NO: 38 which reduce the antigen's similarity withhuman proteins.

sta003

The ‘sta003’ antigen is annotated as ‘surface protein’. In the NCTC 8325strain sta003 is SAOUHSC_(—)00400 and has amino acid sequence SEQ ID NO:39 (GI:88194195). In the Newman strain it is nwmn_(—)0401(GI:151220613).

Useful sta003 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 39 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 39; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 39, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta003 proteins include variants of SEQ ID NO:39. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 39.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 39 while retaining at least oneepitope of SEQ ID NO: 39. The first 32 N-terminal amino acids of SEQ IDNO: 39 can usefully be omitted. Other fragments omit one or more proteindomains.

sta004

The ‘sta004’ antigen is annotated as ‘Siderophore binding protein FatB’.In the NCTC 8325 strain sta004 is SAOUHSC_(—)00749 and has amino acidsequence SEQ ID NO: 40 (GI:88194514). In the Newman strain it isnwmn_(—)0705 (GI:151220917).

Useful sta004 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 40 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 40; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 40, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta004 proteins include variants of SEQ ID NO:40. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 40.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 40 while retaining at least oneepitope of SEQ ID NO: 40. The first 18 N-terminal amino acids of SEQ IDNO: 40 can usefully be omitted. Other fragments omit one or more proteindomains.

sta005

The ‘sta005’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta005 is SAOUHSC_(—)01127 and has amino acid sequenceSEQ ID NO: 41 (GI:88194870). In the Newman strain it is nwmn_(—)1077(GI:151221289).

Useful sta005 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 41 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 41; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 41, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta005 proteins include variants of SEQ ID NO: 41.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 41. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 41 while retaining at least oneepitope of SEQ ID NO: 41. The first 18 N-terminal amino acids of SEQ IDNO: 41 can usefully be omitted. Other fragments omit one or more proteindomains.

sta006

The ‘sta006’ antigen is annotated as ‘ferrichrome-binding protein’, andhas also been referred to as ‘FhuD2’ in the literature [72]. In the NCTC8325 strain sta006 is SAOUHSC_(—)02554 and has amino acid sequence SEQID NO: 42 (GI:88196199). In the Newman strain it is nwmn_(—)2185(GI:151222397).

Useful sta006 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 42 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 42; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 42, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta006 proteins include variants of SEQ ID NO:42. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 42.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 42 while retaining at least oneepitope of SEQ ID NO: 42. The first 17 N-terminal amino acids of SEQ IDNO: 42 can usefully be omitted (to provide SEQ ID NO: 246). Otherfragments omit one or more protein domains. Mutant forms of sta006 arereported in reference 73. A sta006 antigen may be lipidated e.g. with anacylated N-terminus cysteine. One useful sta006 sequence is SEQ ID NO:248, which has a Met-Ala-Ser- sequence at the N-terminus.

sta007

The ‘sta007’ antigen is annotated as ‘secretory antigen precursor’. Inthe NCTC 8325 strain sta007 is SAOUHSC_(—)02571 and has amino acidsequence SEQ ID NO: 43 (GI:88196215). In the Newman strain it isnwmn_(—)2199 (GI:151222411). Proteomic analysis has revealed that thisprotein is secreted or surface-exposed.

Useful sta007 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 43 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 43; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 43, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta007 proteins include variants of SEQ ID NO:43. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 43.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 43 while retaining at least oneepitope of SEQ ID NO: 43. The first 27 N-terminal amino acids of SEQ IDNO: 43 can usefully be omitted. Other fragments omit one or more proteindomains.

sta008

The ‘sta008’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta008 is SAOUHSC_(—)02650 and has amino acid sequence SEQ ID NO:44 (GI:88196290). In the Newman strain it is nwmn_(—)2270(GI:151222482).

Useful sta008 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 44 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 44; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 44, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta008 proteins include variants of SEQ ID NO: 44.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 44. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 44 while retaining at least oneepitope of SEQ ID NO: 44. The first 17 N-terminal amino acids of SEQ IDNO: 44 can usefully be omitted. Other fragments omit one or more proteindomains.

sta009

The ‘sta009’ antigen is annotated as ‘immunoglobulin G-binding proteinSbi’. In the NCTC 8325 strain sta009 is SAOUHSC_(—)02706 and has aminoacid sequence SEQ ID NO: 45 (GI:88196346). In the Newman strain it isnwmn_(—)2317 (GI:151222529).

Useful sta009 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 45 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 45; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 45, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta009 proteins include variants of SEQ ID NO:45. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 45.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 45 while retaining at least oneepitope of SEQ ID NO: 45. The first 29 N-terminal amino acids of SEQ IDNO: 45 can usefully be omitted. Other fragments omit one or more proteindomains.

sta010

The ‘sta010’ antigen is annotated as ‘immunodominant antigen A’. In theNCTC 8325 strain sta010 is SAOUHSC_(—)02887 and has amino acid sequenceSEQ ID NO: 46 (GI:88196515). In the Newman strain it is nwmn_(—)2469(GI:151222681). Proteomic analysis has revealed that this protein issecreted or surface-exposed.

Useful sta010 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 46 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 46; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 46, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta010 proteins include variants of SEQ ID NO: 46.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 46. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 46 while retaining at least oneepitope of SEQ ID NO: 46. The first 29 N-terminal amino acids of SEQ IDNO: 46 can usefully be omitted. Other fragments omit one or more proteindomains.

sta011

The ‘sta011’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta011 is SAOUHSC_(—)00052 and has amino acid sequence SEQ ID NO:47 (GI:88193872).

Useful sta011 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 47 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 47; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 47, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta011 proteins include variants of SEQ ID NO:47. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 47.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 47 while retaining at least oneepitope of SEQ ID NO: 47. The first 23 N-terminal amino acids of SEQ IDNO: 47 can usefully be omitted (to provide SEQ ID NO: 247). Otherfragments omit one or more protein domains. A sta011 antigen may belipidated e.g. with an acylated N-terminus cysteine. One useful sta011sequence is SEQ ID NO: 249, which has a N-terminus methionine.

Variant forms of SEQ ID NO: 47 which may be used as or for preparingsta011 antigens include, but are not limited to, SEQ ID NOs: 213, 214and 215 with various Ile/Val/Leu substitutions.

Sta011 can exist as a monomer or an oligomer, with Ca⁺⁺ ions favouringoligomerisation. The invention can use monomers and/or oligomers ofSta011.

sta012

The ‘sta012’ antigen is annotated as ‘protein with leader’. In the NCTC8325 strain sta012 is SAOUHSC_(—)00106 and has amino acid sequence SEQID NO: 48 (GI:88193919).

Useful sta012 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 48 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 48; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 48, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta012 proteins include variants of SEQ ID NO:48. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 48.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 48 while retaining at least oneepitope of SEQ ID NO: 48. The first 21 N-terminal amino acids of SEQ IDNO: 48 can usefully be omitted. Other fragments omit one or more proteindomains.

sta013

The ‘sta013’ antigen is annotated as ‘poly-gamma-glutamate capsulebiosynthesis protein’. In the NCTC 8325 strain sta013 isSAOUHSC_(—)00107 and has amino acid sequence SEQ ID NO: 49(GI:88193920).

Useful sta013 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 49 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 49; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 49, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta013 proteins include variants of SEQ ID NO:49. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 49.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 49 while retaining at least oneepitope of SEQ ID NO: 49. Other fragments omit one or more proteindomains.

sta014

The ‘sta014’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta014 is SAOUHSC_(—)00137 and has amino acid sequence SEQ ID NO:50 (GI:88193950).

Useful sta014 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 50 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 50; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 50, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta014 proteins include variants of SEQ ID NO:50. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 50.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 50 while retaining at least oneepitope of SEQ ID NO: 50. The first 17 N-terminal amino acids of SEQ IDNO: 50 can usefully be omitted. Other fragments omit one or more proteindomains.

sta015

The ‘sta015’ antigen is annotated as ‘extracellular solute-bindingprotein; RGD containing lipoprotein’. In the NCTC 8325 strain sta015 isSAOUHSC_(—)00170 and has amino acid sequence SEQ ID NO: 51(GI:88193980).

Useful sta015 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 51 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 51; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 51, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta015 proteins include variants of SEQ ID NO:51. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 51.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 51 while retaining at least oneepitope of SEQ ID NO: 51. The first 18 N-terminal amino acids of SEQ IDNO: 51 can usefully be omitted. Other fragments omit one or more proteindomains.

sta016

The ‘sta016’ antigen is annotated as ‘gamma-glutamyltranspeptidase’. Inthe NCTC 8325 strain sta016 is SAOUHSC_(—)00171 and has amino acidsequence SEQ ID NO: 52 (GI:88193981).

Useful sta016 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 52 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 52; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 52, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta016 proteins include variants of SEQ ID NO:52. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 52.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 52 while retaining at least oneepitope of SEQ ID NO: 52. Other fragments omit one or more proteindomains.

sta017

The ‘sta017’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta017 is SAOUHSC_(—)00186 and has amino acid sequence SEQ ID NO:53 (GI:88193996).

Useful sta017 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 53 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 53; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 53, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta017 proteins include variants of SEQ ID NO:53. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 53.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 53 while retaining at least oneepitope of SEQ ID NO: 53. The first 17 N-terminal amino acids of SEQ IDNO: 53 can usefully be omitted. Other fragments omit one or more proteindomains.

sta018

The ‘sta018’ antigen is annotated as ‘extracellular solute-bindingprotein’. In the NCTC 8325 strain sta018 is SAOUHSC_(—)00201 and hasamino acid sequence SEQ ID NO: 54 (GI:88194011).

Useful sta018 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 54 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 54; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 54, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta018 proteins include variants of SEQ ID NO:54. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 54.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 54 while retaining at least oneepitope of SEQ ID NO: 54. Other fragments omit one or more proteindomains.

sta019

The ‘sta019’ antigen is annotated as ‘peptidoglycan hydrolase’. In theNCTC 8325 strain sta019 is SAOUHSC_(—)00248 and has amino acid sequenceSEQ ID NO: 55 (GI:88194055). In the Newman strain it is nwmn_(—)0210(GI:151220422).

Useful sta019 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 55 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 55; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 55, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta019 proteins include variants of SEQ ID NO:55. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 55.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 55 while retaining at least oneepitope of SEQ ID NO: 55. The first 25 N-terminal amino acids of SEQ IDNO: 55 can usefully be omitted. Other fragments omit one or more proteindomains. Useful fragments are SEQ ID NOs: 228 and 229.

Sta019 does not adsorb well to aluminium hydroxide adjuvants, so Sta019present in a composition may me unadsorbed or may be adsorbed to analternative adjuvant e.g. to an aluminium phosphate.

sta020

The ‘sta020’ antigen is annotated as ‘exported protein’. In the NCTC8325 strain sta020 is SAOUHSC_(—)00253 and has amino acid sequence SEQID NO: 56 (GI:88194059).

Useful sta020 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 56 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 56; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 56, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta020 proteins include variants of SEQ ID NO:56. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 56.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 56 while retaining at least oneepitope of SEQ ID NO: 56. The first 30 N-terminal amino acids of SEQ IDNO: 56 can usefully be omitted. Other fragments omit one or more proteindomains.

sta021

The ‘sta021’ antigen is annotated as ‘secretory antigen SsaA-likeprotein’. In the NCTC 8325 strain sta021 is SAOUHSC_(—)00256 and hasamino acid sequence SEQ ID NO: 57 (GI:88194062).

Useful sta021 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 57 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 57; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 57, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta021 proteins include variants of SEQ ID NO:57. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 57.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 57 while retaining at least oneepitope of SEQ ID NO: 57. The first 24 N-terminal amino acids of SEQ IDNO: 57 can usefully be omitted. Other fragments omit one or more proteindomains.

sta022

The ‘sta022’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta022 is SAOUHSC_(—)00279 and has amino acid sequence SEQ ID NO:58 (GI:88194083).

Useful sta022 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 58 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 58; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 58, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 ormore). These sta022 proteins include variants of SEQ ID NO: 58.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 58. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 58 while retaining at least oneepitope of SEQ ID NO: 58. The first 17 N-terminal amino acids of SEQ IDNO: 58 can usefully be omitted. Other fragments omit one or more proteindomains.

sta023

The ‘sta023’ antigen is annotated as ‘5’-nucleotidase; lipoprotein e(P4)family′. In the NCTC 8325 strain sta023 is SAOUHSC_(—)00284 and hasamino acid sequence SEQ ID NO: 59 (GI:88194087).

Useful sta023 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 59 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 59; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 59, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta023 proteins include variants of SEQ ID NO:59. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 59.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 59 while retaining at least oneepitope of SEQ ID NO: 59. The first 31 N-terminal amino acids of SEQ IDNO: 59 can usefully be omitted. Other fragments omit one or more proteindomains.

sta024

The ‘sta024’ antigen is annotated as ‘lipase precursor’. In the NCTC8325 strain sta024 is SAOUHSC_(—)00300 and has amino acid sequence SEQID NO: 60 (GI:88194101).

Useful sta024 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 60 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 60; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 60, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta024 proteins include variants of SEQ ID NO:60. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 60.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 60 while retaining at least oneepitope of SEQ ID NO: 60. The first 37 N-terminal amino acids of SEQ IDNO: 60 can usefully be omitted. Other fragments omit one or more proteindomains.

sta025

The ‘sta025’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta025 is SAOUHSC_(—)00362 and has amino acid sequence SEQ ID NO:61 (GI:88194160).

Useful sta025 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 61 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 61; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 61, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta025 proteins include variants of SEQ ID NO: 61.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 61. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 61 while retaining at least oneepitope of SEQ ID NO: 61. The first 19 N-terminal amino acids of SEQ IDNO: 61 can usefully be omitted. Other fragments omit one or more proteindomains.

sta026

The ‘sta026’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta026 is SAOUHSC_(—)00404 and has amino acid sequence SEQ ID NO:62 (GI:88194198).

Useful sta026 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 62 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 62; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 62, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta026 proteins include variants of SEQ ID NO:62. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 62.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 62 while retaining at least oneepitope of SEQ ID NO: 62. The first 22 N-terminal amino acids of SEQ IDNO: 62 can usefully be omitted. Other fragments omit one or more proteindomains.

sta027

The ‘sta027’ antigen is annotated as ‘probable lipase’. In the NCTC 8325strain sta027 is SAOUHSC_(—)00661 and has amino acid sequence SEQ ID NO:63 (GI:88194426).

Useful sta027 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 63 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 63; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 63, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta027 proteins include variants of SEQ ID NO:63. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 63.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 63 while retaining at least oneepitope of SEQ ID NO: 63. The first 23 N-terminal amino acids of SEQ IDNO: 63 can usefully be omitted. Other fragments omit one or more proteindomains.

sta028

The ‘sta028’ antigen is annotated as ‘secretory antigen SsaA-likeprotein’. In the NCTC 8325 strain sta028 is SAOUHSC_(—)00671 and hasamino acid sequence SEQ ID NO: 64 (GI:88194436). In the Newman strain itis nwmn_(—)0634 (GI:151220846).

Useful sta028 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 64 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 64; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 64, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta028 proteins include variants of SEQ ID NO:64. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 64.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 64 while retaining at least oneepitope of SEQ ID NO: 64. The first 25 N-terminal amino acids of SEQ IDNO: 64 can usefully be omitted. Other fragments omit one or more proteindomains.

sta029

The ‘sta029’ antigen is annotated as ‘ferrichrome binding protein’. Inthe NCTC 8325 strain sta029 is SAOUHSC_(—)00754 and has amino acidsequence SEQ ID NO: 65 (GI:88194518).

Useful sta029 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 65 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 65; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 65, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta029 proteins include variants of SEQ ID NO:65. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 65.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 65 while retaining at least oneepitope of SEQ ID NO: 65. The final 25 C-terminal amino acids of SEQ IDNO: 65 can usefully be omitted. The first 19 N-terminal amino acids ofSEQ ID NO: 65 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta030

The ‘sta030’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta030 is SAOUHSC_(—)00808 and has amino acid sequence SEQ ID NO:66 (GI:88194568).

Useful sta030 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 66 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 66; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 66, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta030 proteins include variants of SEQ ID NO: 66.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 66. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 66 while retaining at least oneepitope of SEQ ID NO: 66. The first 17 N-terminal amino acids of SEQ IDNO: 66 can usefully be omitted. Other fragments omit one or more proteindomains.

sta031

The ‘sta031’ antigen is annotated as ‘5-nucleotidase family protein’. Inthe NCTC 8325 strain sta031 is SAOUHSC_(—)00860 and has amino acidsequence SEQ ID NO: 67 (GI:88194617).

Useful sta031 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 67 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 67; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 67, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta031 proteins include variants of SEQ ID NO:67. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 67.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 67 while retaining at least oneepitope of SEQ ID NO: 67. Other fragments omit one or more proteindomains.

sta032

The ‘sta032’ antigen is annotated as ‘serine protease HtrA’. In the NCTC8325 strain sta032 is SAOUHSC_(—)00958 and has amino acid sequence SEQID NO: 68 (GI:88194715).

Useful sta032 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 68 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 68; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 68, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta032 proteins include variants of SEQ ID NO:68. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 68.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 68 while retaining at least oneepitope of SEQ ID NO: 68. Other fragments omit one or more proteindomains.

sta033

The ‘sta033’ antigen is annotated as ‘cysteine protease precursor’. Inthe NCTC 8325 strain sta033 is SAOUHSC_(—)00987 and has amino acidsequence SEQ ID NO: 69 (GI:88194744).

Useful sta033 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 69 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 69; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 69, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta033 proteins include variants of SEQ ID NO:69. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 69.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 69 while retaining at least oneepitope of SEQ ID NO: 69. The first 29 N-terminal amino acids of SEQ IDNO: 69 can usefully be omitted. Other fragments omit one or more proteindomains.

sta034

The ‘sta034’ antigen is annotated as ‘glutamyl endopeptidase precursor’.In the NCTC 8325 strain sta034 is SAOUHSC_(—)00988 and has amino acidsequence SEQ ID NO: 70 (GI:88194745).

Useful sta034 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 70 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 70; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 70, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta034 proteins include variants of SEQ ID NO:70. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 70.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 70 while retaining at least oneepitope of SEQ ID NO: 70. The first 29 N-terminal amino acids of SEQ IDNO: 70 can usefully be omitted. Other fragments omit one or more proteindomains.

sta035

The ‘sta035’ antigen is annotated as ‘first protein’. In the NCTC 8325strain sta035 is SAOUHSC_(—)00998 and has amino acid sequence SEQ ID NO:71 (GI:88194754).

Useful sta035 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 71 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 71; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 71, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta035 proteins include variants of SEQ ID NO:71. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 71.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 71 while retaining at least oneepitope of SEQ ID NO: 71. The first 25 N-terminal amino acids of SEQ IDNO: 71 can usefully be omitted. Other fragments omit one or more proteindomains.

sta036

The ‘sta036’ antigen is annotated as ‘iron-regulated protein withleader’. In the NCTC 8325 strain sta036 is SAOUHSC_(—)01084 and hasamino acid sequence SEQ ID NO: 72 (GI:88194831).

Useful sta036 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 72 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 72; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 72, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta036 proteins include variants of SEQ ID NO:72. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 72.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 72 while retaining at least oneepitope of SEQ ID NO: 72. The final 27 C-terminal amino acids of SEQ IDNO: 72 can usefully be omitted. The first 32 N-terminal amino acids ofSEQ ID NO: 72 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta037

The ‘sta037’ antigen is annotated as ‘iron ABC transporter; iron-bindingprotein IsdE’. In the NCTC 8325 strain sta037 is SAOUHSC_(—)01085 andhas amino acid sequence SEQ ID NO: 73 (GI: 88194832).

Useful sta037 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 73 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 73; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 73, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta037 proteins include variants of SEQ ID NO:73. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 73.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 73 while retaining at least oneepitope of SEQ ID NO: 73. The first 9 N-terminal amino acids of SEQ IDNO: 73 can usefully be omitted. Other fragments omit one or more proteindomains.

sta038

The ‘sta038’ antigen is annotated as ‘NPQTN specific sortase B’. In theNCTC 8325 strain sta038 is SAOUHSC_(—)01088 and has amino acid sequenceSEQ ID NO: 74 (GI:88194835).

Useful sta038 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 74 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 74; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 74, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta038 proteins include variants of SEQ ID NO: 74.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 74. Otherpreferred fragments lack one or more amino acids, (e.g. 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 74 while retaining at least oneepitope of SEQ ID NO: 74. The first 21 N-terminal amino acids of SEQ IDNO: 74 can usefully be omitted. Other fragments omit one or more proteindomains.

sta039

The ‘sta039’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta039 is SAOUHSC_(—)01124 and has amino acid sequenceSEQ ID NO: 75 (GI:88194868).

Useful sta039 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 75 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% more) toSEQ ID NO: 75; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 75, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta039 proteins include variants of SEQ ID NO: 75.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 75. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 75 while retaining at least oneepitope of SEQ ID NO: 75. The first 22 N-terminal amino acids of SEQ IDNO: 75 can usefully be omitted. Other fragments omit one or more proteindomains.

sta040

The ‘sta040’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta040 is SAOUHSC_(—)01125 and has amino acid sequenceSEQ ID NO: 76 (GI:88194869). In the Newman strain it is nwmn_(—)1076(GI:151221288).

Useful sta040 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 76 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 76; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 76, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta040 proteins include variants of SEQ ID NO: 76.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 76. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 76 while retaining at least oneepitope of SEQ ID NO: 76. The first 21 N-terminal amino acids of SEQ IDNO: 76 can usefully be omitted. Other fragments omit one or more proteindomains.

sta041

The ‘sta041’ antigen is annotated as ‘fibronectin-binding proteinA-related’. In the NCTC 8325 strain sta041 is SAOUHSC_(—)01175 and hasamino acid sequence SEQ ID NO: 77 (GI:88194914).

Useful sta041 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 77 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 77; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 77, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta041 proteins include variants of SEQ ID NO:77. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 77.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 77 while retaining at least oneepitope of SEQ ID NO: 77. Other fragments omit one or more proteindomains.

sta042

The ‘sta042’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta042 is SAOUHSC_(—)01180 and has amino acid sequence SEQ ID NO:78 (GI:88194919).

Useful sta042 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 78 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 78; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 78, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta042 proteins include variants of SEQ ID NO:78. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 78.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 78 while retaining at least oneepitope of SEQ ID NO: 78. The first 18 N-terminal amino acids of SEQ IDNO: 78 can usefully be omitted. Other fragments omit one or more proteindomains.

sta043

The ‘sta043’ antigen is annotated as ‘cell wall hydrolase’. In the NCTC8325 strain sta043 is SAOUHSC_(—)01219 and has amino acid sequence SEQID NO: 79 (GI:88194955).

Useful sta043 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 79 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 79; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 79, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta043 proteins include variants of SEQ ID NO:79. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 79.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 79 while retaining at least oneepitope of SEQ ID NO: 79. The first 38 N-terminal amino acids of SEQ IDNO: 79 can usefully be omitted. Other fragments omit one or more proteindomains.

sta044

The ‘sta044’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta044 is SAOUHSC_(—)01508 and has amino acid sequence SEQ ID NO:80 (GI:88195223).

Useful sta044 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 80 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 80; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 80, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta044 proteins include variants of SEQ ID NO:80. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 80.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 80 while retaining at least oneepitope of SEQ ID NO: 80. The first 17 N-terminal amino acids of SEQ IDNO: 80 can usefully be omitted. Other fragments omit one or more proteindomains.

sta045

The ‘sta045’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta045 is SAOUHSC_(—)01627 and has amino acid sequence SEQ ID NO:81 (GI:88195337).

Useful sta045 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 81 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 81; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 81, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These sta045 proteins include variants of SEQ ID NO: 81.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 81. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 81 while retaining at least oneepitope of SEQ ID NO: 81. The first 16 N-terminal amino acids of SEQ IDNO: 81 can usefully be omitted. Other fragments omit one or more proteindomains.

sta046

The ‘sta046’ antigen is annotated as ‘Excalibur protein’. In the NCTC8325 strain sta046 is SAOUHSC_(—)01918 and has amino acid sequence SEQID NO: 82 (GI:88195613).

Useful sta046 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 82 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 82; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 82, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta046 proteins include variants of SEQ ID NO: 82.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 82. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 82 while retaining at least oneepitope of SEQ ID NO: 82. The first 53 N-terminal amino acids of SEQ IDNO: 82 can usefully be omitted. Other fragments omit one or more proteindomains.

sta047

The ‘sta047’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta047 is SAOUHSC_(—)01920 and has amino acid sequence SEQ ID NO:83 (GI:88195615).

Useful sta047 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 83 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 83; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 83, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta047 proteins include variants of SEQ ID NO: 83.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 83. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 83 while retaining at least oneepitope of SEQ ID NO: 83. The first 18 N-terminal amino acids of SEQ IDNO: 83 can usefully be omitted. Other fragments omit one or more proteindomains.

sta048

The ‘sta048’ antigen is annotated as ‘intracellular serine protease’. Inthe NCTC 8325 strain sta048 is SAOUHSC_(—)01949 and has amino acidsequence SEQ ID NO: 84 (GI:88195642).

Useful sta048 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 84 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 84; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 84, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta048 proteins include variants of SEQ ID NO:84. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 84.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 84 while retaining at least oneepitope of SEQ ID NO: 84. The first 27 N-terminal amino acids of SEQ IDNO: 84 can usefully be omitted. Other fragments omit one or more proteindomains.

sta049

The ‘sta049’ antigen is annotated as ‘protein export protein PrsA’. Inthe NCTC 8325 strain sta049 is SAOUHSC_(—)01972 and has amino acidsequence SEQ ID NO: 85 (GI:88195663). In the Newman strain it isnwmn_(—)1733 (GI:151221945).

Useful sta049 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 85 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 85; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 85, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta049 proteins include variants of SEQ ID NO:85. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 85.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 85 while retaining at least oneepitope of SEQ ID NO: 85. The first 25 N-terminal amino acids of SEQ IDNO: 85 can usefully be omitted. Other fragments omit one or more proteindomains.

sta050

The ‘sta050’ antigen is annotated as ‘staphopain thiol proteinase’. Inthe NCTC 8325 strain sta050 is SAOUHSC_(—)02127 and has amino acidsequence SEQ ID NO: 86 (GI:88195808).

Useful sta050 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 86 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 86; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 86, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta050 proteins include variants of SEQ ID NO:86. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 86.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 86 while retaining at least oneepitope of SEQ ID NO: 86. The first 25 N-terminal amino acids of SEQ IDNO: 86 can usefully be omitted. Other fragments omit one or more proteindomains.

sta051

The ‘sta051’ antigen is annotated as ‘protein with leader’. In the NCTC8325 strain sta051 is SAOUHSC_(—)02147 and has amino acid sequence SEQID NO: 87 (GI:88195827).

Useful sta051 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 87 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 87; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 87, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta051 proteins include variants of SEQ ID NO:87. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 87.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 87 while retaining at least oneepitope of SEQ ID NO: 87. The first 24 N-terminal amino acids of SEQ IDNO: 87 can usefully be omitted. Other fragments omit one or more proteindomains.

sta052

The ‘sta052’ antigen is annotated as ‘ferric hydroxamate receptor 1’. Inthe NCTC 8325 strain sta052 is SAOUHSC_(—)02246 and has amino acidsequence SEQ ID NO: 88 (GI:88195918).

Useful sta052 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 88 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 88; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 88, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta052 proteins include variants of SEQ ID NO:88. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 88.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 88 while retaining at least oneepitope of SEQ ID NO: 88. The first 17 N-terminal amino acids of SEQ IDNO: 88 can usefully be omitted. Other fragments omit one or more proteindomains.

sta053

The ‘sta053’ antigen is annotated as ‘srdH family protein’. In the NCTC8325 strain sta053 is SAOUHSC_(—)02257 and has amino acid sequence SEQID NO: 89 (GI:88195928).

Useful sta053 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 89 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 89; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 89, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta053 proteins include variants of SEQ ID NO:89. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 89.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 89 while retaining at least oneepitope of SEQ ID NO: 89. The first 26 N-terminal amino acids of SEQ IDNO: 89 can usefully be omitted. Other fragments omit one or more proteindomains.

sta054

The ‘sta054’ antigen is annotated as ‘Probable transglycosylase isaAprecursor’. In the NCTC 8325 strain sta054 is SAOUHSC_(—)02333 and hasamino acid sequence SEQ ID NO: 90 (GI:88195999).

Useful sta054 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 90 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 90; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 90, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200 or more). These sta054 proteins include variants of SEQ ID NO: 90.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 90. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 90 while retaining at least oneepitope of SEQ ID NO: 90. The first 27 N-terminal amino acids of SEQ IDNO: 90 can usefully be omitted. Other fragments omit one or more proteindomains.

sta055

The ‘sta055’ antigen is annotated as ‘surface hydrolase’. In the NCTC8325 strain sta055 is SAOUHSC_(—)02448 and has amino acid sequence SEQID NO: 91 (GI:88196100).

Useful sta055 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 91 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 91; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 91, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta055 proteins include variants of SEQ ID NO:91. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 91.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 91 while retaining at least oneepitope of SEQ ID NO: 91. The first 31 N-terminal amino acids of SEQ IDNO: 91 can usefully be omitted. Other fragments omit one or more proteindomains.

sta056

The ‘sta056’ antigen is annotated as ‘hyaluronate lyase’. In the NCTC8325 strain sta056 is SAOUHSC_(—)02463 and has amino acid sequence SEQID NO: 92 (GI:88196115).

Useful sta056 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 92 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 92; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 92, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta056 proteins include variants of SEQ ID NO:92. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 92.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 92 while retaining at least oneepitope of SEQ ID NO: 92. The first 24 N-terminal amino acids of SEQ IDNO: 92 can usefully be omitted. Other fragments omit one or more proteindomains.

sta057

The ‘sta057’ antigen is annotated as ‘secretory antigen precursor SsaA’.In the NCTC 8325 strain sta057 is SAOUHSC_(—)02576 and has amino acidsequence SEQ ID NO: 93 (GI:88196220). In the Newman strain it isnwmn_(—)2203 (GI:151222415).

Useful sta057 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 93 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 93; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 93, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150or more). These sta057 proteins include variants of SEQ ID NO: 93.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 93. Otherpreferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one or moreamino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more)from the N-terminus of SEQ ID NO: 93 while retaining at least oneepitope of SEQ ID NO: 93. The first 27 N-terminal amino acids of SEQ IDNO: 93 can usefully be omitted. Other fragments omit one or more proteindomains.

sta058

The ‘sta058’ antigen is annotated as ‘Zn-binding lipoprotein adcA-like’.In the NCTC 8325 strain sta058 is SAOUHSC_(—)02690 and has amino acidsequence SEQ ID NO: 94 (GI:88196330).

Useful sta058 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 94 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 94; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 94, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta058 proteins include variants of SEQ ID NO:94. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 94.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 94 while retaining at least oneepitope of SEQ ID NO: 94. The first 20 N-terminal amino acids of SEQ IDNO: 94 can usefully be omitted. Other fragments omit one or more proteindomains.

sta059

The ‘sta059’ antigen is annotated as ‘gamma-hemolysin h-gamma-iisubunit’. In the NCTC 8325 strain sta059 is SAOUHSC_(—)02708 and hasamino acid sequence SEQ ID NO: 95 (GI:88196348).

Useful sta059 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 95 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 95; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 95, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta059 proteins include variants of SEQ ID NO:95. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 95.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 95 while retaining at least oneepitope of SEQ ID NO: 95. The first 20 N-terminal amino acids of SEQ IDNO: 95 can usefully be omitted. Other fragments omit one or more proteindomains.

sta060

The ‘sta060’ antigen is annotated as ‘peptide ABC transporter;peptide-binding protein’. In the NCTC 8325 strain sta060 isSAOUHSC_(—)02767 and has amino acid sequence SEQ ID NO: 96(GI:88196403).

Useful sta060 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 96 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 96; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 96, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta060 proteins include variants of SEQ ID NO:96. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 96.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 96 while retaining at least oneepitope of SEQ ID NO: 96. The first 20 N-terminal amino acids of SEQ IDNO: 96 can usefully be omitted. Other fragments omit one or more proteindomains.

sta061

The ‘sta061’ antigen is annotated as ‘protein with leader’. In the NCTC8325 strain sta061 is SAOUHSC_(—)02783 and has amino acid sequence SEQID NO: 97 (GI:88196419).

Useful sta061 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 97 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 97; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 97, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta061 proteins include variants of SEQ ID NO:97. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 97.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 97 while retaining at least oneepitope of SEQ ID NO: 97. The first 21 N-terminal amino acids of SEQ IDNO: 97 can usefully be omitted. Other fragments omit one or more proteindomains.

sta062

The ‘sta062’ antigen is annotated as ‘protein with leader’. In the NCTC8325 strain sta062 is SAOUHSC_(—)02788 and has amino acid sequence SEQID NO: 98 (GI:88196424).

Useful sta062 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 98 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 98; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 98, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta062 proteins include variants of SEQ ID NO:98. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 98.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 98 while retaining at least oneepitope of SEQ ID NO: 98. The first 22 N-terminal amino acids of SEQ IDNO: 98 can usefully be omitted. Other fragments omit one or more proteindomains.

sta063

The ‘sta063’ antigen is annotated as ‘aureolysin’. In the NCTC 8325strain sta063 is SAOUHSC_(—)02971 and has amino acid sequence SEQ ID NO:99 (GI:88196592).

Useful sta063 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 99 and/or may comprise an amino acidsequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%,85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) toSEQ ID NO: 99; and/or (b) comprising a fragment of at least ‘n’consecutive amino acids of SEQ ID NO: 99, wherein ‘n’ is 7 or more (e.g.8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 150,200, 250 or more). These sta063 proteins include variants of SEQ ID NO:99. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 99.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 99 while retaining at least oneepitope of SEQ ID NO: 99. The first 16 N-terminal amino acids of SEQ IDNO: 99 can usefully be omitted. Other fragments omit one or more proteindomains.

sta064

The ‘sta064’ antigen is annotated as ‘lipase’. In the NCTC 8325 strainsta064 is SAOUHSC_(—)03006 and has amino acid sequence SEQ ID NO: 100(GI:88196625). In the Newman strain it is nwmn_(—)2569 (GI:151222781).

Useful sta064 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 100 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 100; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 100, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta064 proteins include variants ofSEQ ID NO: 100. Preferred fragments of (b) comprise an epitope from SEQID NO: 100. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 100 while retaining atleast one epitope of SEQ ID NO: 100. The first 34 N-terminal amino acidsof SEQ ID NO: 100 can usefully be omitted. Other fragments omit one ormore protein domains.

sta065

The ‘sta065’ antigen is annotated as ‘1-phosphatidylinositolphosphodiesterase precursor’. In the NCTC 8325 strain sta065 isSAOUHSC_(—)00051 and has amino acid sequence SEQ ID NO: 101(GI:88193871).

Useful sta065 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 101 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 101; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 101, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta065 proteins include variants ofSEQ ID NO: 101. Preferred fragments of (b) comprise an epitope from SEQID NO: 101. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 101 while retaining atleast one epitope of SEQ ID NO: 101. The first 26 N-terminal amino acidsof SEQ ID NO: 101 can usefully be omitted. Other fragments omit one ormore protein domains.

sta066

The ‘sta066’ antigen is annotated as ‘protein’. In the NCTC 8325 strainsta066 is SAOUHSC_(—)00172 and has amino acid sequence SEQ ID NO: 102(GI:88193982).

Useful sta066 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 102 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 102; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 102, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta066 proteins include variants ofSEQ ID NO: 102. Preferred fragments of (b) comprise an epitope from SEQID NO: 102. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 102 while retaining atleast one epitope of SEQ ID NO: 102. The first 21 N-terminal amino acidsof SEQ ID NO: 102 can usefully be omitted. Other fragments omit one ormore protein domains.

sta067

The ‘sta067’ antigen is annotated as ‘bacterial extracellularsolute-binding protein’. In the NCTC 8325 strain sta067 isSAOUHSC_(—)00176 and has amino acid sequence SEQ ID NO: 103(GI:88193986).

Useful sta067 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 103 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 103; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 103, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta067 proteins include variants ofSEQ ID NO: 103. Preferred fragments of (b) comprise an epitope from SEQID NO: 103. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 103 while retaining atleast one epitope of SEQ ID NO: 103. The first 20 N-terminal amino acidsof SEQ ID NO: 103 can usefully be omitted. Other fragments omit one ormore protein domains.

sta068

The ‘sta068’ antigen is annotated as ‘iron permease FTR1’. In the NCTC8325 strain sta068 is SAOUHSC_(—)00327 and has amino acid sequence SEQID NO: 104 (GI:88194127).

Useful sta068 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 104 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 104; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 104, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta068 proteins include variants ofSEQ ID NO: 104. Preferred fragments of (b) comprise an epitope from SEQID NO: 104. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 104 while retaining atleast one epitope of SEQ ID NO: 104. The final 20 C-terminal amino acidsof SEQ ID NO: 104 can usefully be omitted. The first 14 N-terminal aminoacids of SEQ ID NO: 104 can usefully be omitted. Other fragments omitone or more protein domains.

sta069

The ‘sta069’ antigen is annotated as ‘autolysin precursor’. In the NCTC8325 strain sta069 is SAOUHSC_(—)00427 and has amino acid sequence SEQID NO: 105 (GI:88194219).

Useful sta069 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 105 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 105; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 105, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta069 proteins include variants ofSEQ ID NO: 105. Preferred fragments of (b) comprise an epitope from SEQID NO: 105. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 105 while retaining atleast one epitope of SEQ ID NO: 105. The first 25 N-terminal amino acidsof SEQ ID NO: 105 can usefully be omitted. Other fragments omit one ormore protein domains.

sta070

The ‘sta070’ antigen is annotated as ‘immunogenic secretedprecursor-like protein (truncated)’. In the NCTC 8325 strain sta070 isSAOUHSC_(—)00773 and has amino acid sequence SEQ ID NO: 106(GI:88194535).

Useful sta070 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 106 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 106; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 106, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta070 proteins include variants ofSEQ ID NO: 106. Preferred fragments of (b) comprise an epitope from SEQID NO: 106. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 106 while retaining atleast one epitope of SEQ ID NO: 106. The first 24 N-terminal amino acidsof SEQ ID NO: 106 can usefully be omitted. Other fragments omit one ormore protein domains.

sta071

The ‘sta071’ antigen is annotated as ‘hemolysin’. In the NCTC 8325strain sta071 is SAOUHSC_(—)00854 and has amino acid sequence SEQ ID NO:107 (GI:88194612).

Useful sta071 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 107 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 107; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 107, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta071 proteins include variants ofSEQ ID NO: 107. Preferred fragments of (b) comprise an epitope from SEQID NO: 107. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 107 while retaining atleast one epitope of SEQ ID NO: 107. The first 24 N-terminal amino acidsof SEQ ID NO: 107 can usefully be omitted. Other fragments omit one ormore protein domains.

sta072

The ‘sta072’ antigen is annotated as ‘extramembranal protein’. In theNCTC 8325 strain sta072 is SAOUHSC_(—)00872 and has amino acid sequenceSEQ ID NO: 108 (GI:88194629).

Useful sta072 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 108 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 108; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 108, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta072 proteins include variants ofSEQ ID NO: 108. Preferred fragments of (b) comprise an epitope from SEQID NO: 108. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 108 while retaining atleast one epitope of SEQ ID NO: 108. The first 24 N-terminal amino acidsof SEQ ID NO: 108 can usefully be omitted. Other fragments omit one ormore protein domains.

sta073

The ‘sta073’ antigen is annotated as ‘bifunctional autolysin precursor’.In the NCTC 8325 strain sta073 is SAOUHSC_(—)00994 and has amino acidsequence SEQ ID NO: 109 (GI:88194750). In the Newman strain it isnwmn_(—)0922 (GI:151221134). Proteomic analysis has revealed that thisprotein is secreted or surface-exposed.

Useful sta073 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 109 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 109; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 109, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta073 proteins include variants ofSEQ ID NO: 109. Preferred fragments of (b) comprise an epitope from SEQID NO: 109. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 109 while retaining atleast one epitope of SEQ ID NO: 109. The first 24 N-terminal amino acidsof SEQ ID NO: 109 can usefully be omitted. Other fragments omit one ormore protein domains.

A Sta073 antigen can usefully be included in a composition incombination with a Sta112 [74].

Sta073 does not adsorb well to aluminium hydroxide adjuvants, so Sta073present in a composition may be unadsorbed or may be adsorbed to analternative adjuvant e.g. to an aluminium phosphate.

sta074

The ‘sta074’ antigen is annotated as ‘factor essential for methicillinresistance’. In the NCTC 8325 strain sta074 is SAOUHSC_(—)01220 and hasamino acid sequence SEQ ID NO: 110 (GI:88194956).

Useful sta074 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 110 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 110; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 110, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta074 proteins include variants ofSEQ ID NO: 110. Preferred fragments of (b) comprise an epitope from SEQID NO: 110. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 110 while retaining atleast one epitope of SEQ ID NO: 110. Other fragments omit one or moreprotein domains.

sta075

The ‘sta075’ antigen is annotated as ‘insulysin; peptidase family M16’.In the NCTC 8325 strain sta075 is SAOUHSC_(—)01256 and has amino acidsequence SEQ ID NO: 111 (GI:88194989).

Useful sta075 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 111 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 111; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 111, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta075 proteins include variants ofSEQ ID NO: 111. Preferred fragments of (b) comprise an epitope from SEQID NO: 111. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 111 while retaining atleast one epitope of SEQ ID NO: 111. Other fragments omit one or moreprotein domains.

sta076

The ‘sta076’ antigen is annotated as ‘hydrolase’. In the NCTC 8325strain sta076 is SAOUHSC_(—)01263 and has amino acid sequence SEQ ID NO:112 (GI:88194996).

Useful sta076 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 112 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 112; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 112, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta076 proteins include variants ofSEQ ID NO: 112. Preferred fragments of (b) comprise an epitope from SEQID NO: 112. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 112 while retaining atleast one epitope of SEQ ID NO: 112. The first 24 N-terminal amino acidsof SEQ ID NO: 112 can usefully be omitted. Other fragments omit one ormore protein domains.

sta077

The ‘sta077’ antigen is annotated as ‘protein’. In the NCTC 8325 strainsta077 is SAOUHSC_(—)01317 and has amino acid sequence SEQ ID NO: 113(GI:88195047). Proteomic analysis has revealed that this protein issecreted or surface-exposed.

Useful sta077 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 113 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 113; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 113, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta077 proteins include variants ofSEQ ID NO: 113. Preferred fragments of (b) comprise an epitope from SEQID NO: 113. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 113 while retaining atleast one epitope of SEQ ID NO: 113. The first 20 N-terminal amino acidsof SEQ ID NO: 113 can usefully be omitted. Other fragments omit one ormore protein domains.

sta078

The ‘sta078’ antigen is annotated as ‘FtsK/SpoIIIE family protein’. Inthe NCTC 8325 strain sta078 is SAOUHSC_(—)01857 and has amino acidsequence SEQ ID NO: 114 (GI:88195555).

Useful sta078 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 114 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 114; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 114, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta078 proteins include variants ofSEQ ID NO: 114. Preferred fragments of (b) comprise an epitope from SEQID NO: 114. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 114 while retaining atleast one epitope of SEQ ID NO: 114. Other fragments omit one or moreprotein domains.

sta079

The ‘sta079’ antigen is annotated as ‘serine protease SplF’. In the NCTC8325 strain sta079 is SAOUHSC_(—)01935 and has amino acid sequence SEQID NO: 115 (GI:88195630).

Useful sta079 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 115 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 115; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 115, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta079 proteins include variants of SEQ IDNO: 115. Preferred fragments of (b) comprise an epitope from SEQ ID NO:115. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 115 while retaining at leastone epitope of SEQ ID NO: 115. The first 36 N-terminal amino acids ofSEQ ID NO: 115 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta080

The ‘sta080’ antigen is annotated as ‘serine protease SplE’. In the NCTC8325 strain sta080 is SAOUHSC_(—)01936 and has amino acid sequence SEQID NO: 116 (GI:88195631).

Useful sta080 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 116 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 116; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 116, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta080 proteins include variants of SEQ IDNO: 116. Preferred fragments of (b) comprise an epitope from SEQ ID NO:116. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 116 while retaining at leastone epitope of SEQ ID NO: 116. The first 36 N-terminal amino acids ofSEQ ID NO: 116 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta081

The ‘sta081’ antigen is annotated as ‘serine protease SplD(EC:3.4.21.19)’. In the NCTC 8325 strain sta081 is SAOUHSC_(—)01938 andhas amino acid sequence SEQ ID NO: 170 (GI:88195633).

Useful sta081 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 170 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 170; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 170, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta081 proteins include variants of SEQ IDNO: 170. Preferred fragments of (b) comprise an epitope from SEQ ID NO:170. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 170 while retaining at leastone epitope of SEQ ID NO: 170. The first 36 N-terminal amino acids ofSEQ ID NO: 170 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta082

The ‘sta082’ antigen is annotated as ‘serine protease SplC’. In the NCTC8325 strain sta082 is SAOUHSC_(—)01939 and has amino acid sequence SEQID NO: 117 (GI:88195634).

Useful sta082 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 117 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 117; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 117, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta082 proteins include variants of SEQ IDNO: 117. Preferred fragments of (b) comprise an epitope from SEQ ID NO:117. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 117 while retaining at leastone epitope of SEQ ID NO: 117. The first 36 N-terminal amino acids ofSEQ ID NO: 117 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta083

The ‘sta083’ antigen is annotated as ‘serine protease SplB’. In the NCTC8325 strain sta083 is SAOUHSC_(—)01941 and has amino acid sequence SEQID NO: 118 (GI:88195635).

Useful sta083 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 118 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 118; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 118, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta083 proteins include variants of SEQ IDNO: 118. Preferred fragments of (b) comprise an epitope from SEQ ID NO:118. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 118 while retaining at leastone epitope of SEQ ID NO: 118. The first 36 N-terminal amino acids ofSEQ ID NO: 118 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta084

The ‘sta084’ antigen is annotated as ‘serine protease SplA’. In the NCTC8325 strain sta084 is SAOUHSC_(—)01942 and has amino acid sequence SEQID NO: 119 (GI:88195636).

Useful sta084 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 119 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 119; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 119, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta084 proteins include variants of SEQ IDNO: 119. Preferred fragments of (b) comprise an epitope from SEQ ID NO:119. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 119 while retaining at leastone epitope of SEQ ID NO: 119. The first 35 N-terminal amino acids ofSEQ ID NO: 119 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta085

The ‘sta085’ antigen is annotated as ‘staphylokinase precursor’. In theNCTC 8325 strain sta085 is SAOUHSC_(—)02171 and has amino acid sequenceSEQ ID NO: 120 (GI:88195848).

Useful sta085 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 120 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 120; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 120, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These sta085 proteins include variants of SEQ ID NO:120. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 120.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 120 while retaining at least oneepitope of SEQ ID NO: 120. The first 27 N-terminal amino acids of SEQ IDNO: 120 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta086

The ‘sta086’ antigen is annotated as ‘OxaA-like protein’. In the NCTC8325 strain sta086 is SAOUHSC_(—)02327 and has amino acid sequence SEQID NO: 121 (GI:88195993).

Useful sta086 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 121 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 121; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 121, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta086 proteins include variants ofSEQ ID NO: 121. Preferred fragments of (b) comprise an epitope from SEQID NO: 121. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 121 while retaining atleast one epitope of SEQ ID NO: 121. The first 19 N-terminal amino acidsof SEQ ID NO: 121 can usefully be omitted. Other fragments omit one ormore protein domains.

sta087

The ‘sta087’ antigen is annotated as ‘teicoplanin resistance proteinTcaA’. In the NCTC 8325 strain sta087 is SAOUHSC_(—)02635 and has aminoacid sequence SEQ ID NO: 122 (GI:88196276).

Useful sta087 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 122 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 122; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 122, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta087 proteins include variants ofSEQ ID NO: 122. Preferred fragments of (b) comprise an epitope from SEQID NO: 122. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 122 while retaining atleast one epitope of SEQ ID NO: 122. Other fragments omit one or moreprotein domains.

sta088

The ‘sta088’ antigen is annotated as ‘esterase’. In the NCTC 8325 strainsta088 is SAOUHSC_(—)02844 and has amino acid sequence SEQ ID NO: 123(GI:88196477).

Useful sta088 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 123 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 123; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 123, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta088 proteins include variants ofSEQ ID NO: 123. Preferred fragments of (b) comprise an epitope from SEQID NO: 123. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 123 while retaining atleast one epitope of SEQ ID NO: 123. The first 18 N-terminal amino acidsof SEQ ID NO: 123 can usefully be omitted. Other fragments omit one ormore protein domains.

sta089

The ‘sta089’ antigen is annotated as ‘LysM domain protein’. In the NCTC8325 strain sta089 is SAOUHSC_(—)02855 and has amino acid sequence SEQID NO: 124 (GI:88196486).

Useful sta089 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 124 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 124; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 124, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta089 proteins include variants of SEQ ID NO: 124.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 124.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 124 while retaining at least oneepitope of SEQ ID NO: 124. The first 20 N-terminal amino acids of SEQ IDNO: 124 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta090

The ‘sta090’ antigen is annotated as ‘LysM domain protein’. In the NCTC8325 strain sta090 is SAOUHSC_(—)02883 and has amino acid sequence SEQID NO: 125 (GI:88196512).

Useful sta090 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 125 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 125; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 125, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta090 proteins include variants ofSEQ ID NO: 125. Preferred fragments of (b) comprise an epitope from SEQID NO: 125. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 125 while retaining atleast one epitope of SEQ ID NO: 125. The first 26 N-terminal amino acidsof SEQ ID NO: 125 can usefully be omitted. Other fragments omit one ormore protein domains.

sta091

The ‘sta091’ antigen is annotated as ‘lipoprotein’. In the NCTC 8325strain sta091 is SAOUHSC_(—)00685 and has amino acid sequence SEQ ID NO:126 (GI:88194450).

Useful sta091 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 126 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 126; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 126, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta091 proteins include variants of SEQ ID NO: 126.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 126.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 126 while retaining at least oneepitope of SEQ ID NO: 126. The first 15 N-terminal amino acids of SEQ IDNO: 126 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta092

The ‘sta092’ antigen is annotated as ‘M23/M37 peptidase domain protein’.In the NCTC 8325 strain sta092 is SAOUHSC_(—)00174 and has amino acidsequence SEQ ID NO: 127 (GI:88193984).

Useful sta092 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 127 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 127; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 127, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These sta092 proteins include variants of SEQ ID NO:127. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 127.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 127 while retaining at least oneepitope of SEQ ID NO: 127. The first 25 N-terminal amino acids of SEQ IDNO: 127 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta093

The ‘sta093’ antigen is annotated as ‘protein’. In the NCTC 8325 strainsta093 is SAOUHSC_(—)01854 and has amino acid sequence SEQ ID NO: 128(GI:88195552).

Useful sta093 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 128 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 128; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 128, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta093 proteins include variants ofSEQ ID NO: 128. Preferred fragments of (b) comprise an epitope from SEQID NO: 128. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 128 while retaining atleast one epitope of SEQ ID NO: 128. Other fragments omit one or moreprotein domains.

sta094

The ‘sta094’ antigen is annotated as ‘protein’. In the NCTC 8325 strainsta094 is SAOUHSC_(—)01512 and has amino acid sequence SEQ ID NO: 129(GI:88195226).

Useful sta094 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 129 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 129; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 129, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta094 proteins include variants ofSEQ ID NO: 129. Preferred fragments of (b) comprise an epitope from SEQID NO: 129. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 129 while retaining atleast one epitope of SEQ ID NO: 129. The first 17 N-terminal amino acidsof SEQ ID NO: 129 can usefully be omitted. Other fragments omit one ormore protein domains.

sta095

The ‘sta095’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta095 is SAOUHSC_(—)00383 and has amino acid sequenceSEQ ID NO: 130 (GI:88194180). In the Newman strain it is nwmn_(—)0388(GI:151220600).

Useful sta095 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 130 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 130; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 130, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta095 proteins include variants of SEQ IDNO: 130. Preferred fragments of (b) comprise an epitope from SEQ ID NO:130. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 130 while retaining at leastone epitope of SEQ ID NO: 130. The first 32 N-terminal amino acids ofSEQ ID NO: 130 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta096

The ‘sta096’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta096 is SAOUHSC_(—)00384 and has amino acid sequenceSEQ ID NO: 131 (GI:88194181).

Useful sta096 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 131 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 131; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 131, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta096 proteins include variants of SEQ IDNO: 131. Preferred fragments of (b) comprise an epitope from SEQ ID NO:131. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 131 while retaining at leastone epitope of SEQ ID NO: 131. The first 30 N-terminal amino acids ofSEQ ID NO: 131 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta097

The ‘sta097’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta097 is SAOUHSC_(—)00386 and has amino acid sequenceSEQ ID NO: 132 (GI:88194182).

Useful sta097 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 132 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 132; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 132, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta097 proteins include variants ofSEQ ID NO: 132. Preferred fragments of (b) comprise an epitope from SEQID NO: 132. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 132 while retaining atleast one epitope of SEQ ID NO: 132. The first 30 N-terminal amino acidsof SEQ ID NO: 132 can usefully be omitted. Other fragments omit one ormore protein domains.

sta098

The ‘sta098’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta098 is SAOUHSC_(—)00389 and has amino acid sequenceSEQ ID NO: 133 (GI:88194184). In the Newman strain it is nwmn_(—)0391(GI:151220603).

Useful sta098 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 133 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 133; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 133, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta098 proteins include variants ofSEQ ID NO: 133. Preferred fragments of (b) comprise an epitope from SEQID NO: 133. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 133 while retaining atleast one epitope of SEQ ID NO: 133. The first 30 N-terminal amino acidsof SEQ ID NO: 133 can usefully be omitted. Other fragments omit one ormore protein domains.

sta099

The ‘sta099’ antigen is annotated as ‘superantigen-like protein 5’. Inthe NCTC 8325 strain sta099 is SAOUHSC_(—)00390 and has amino acidsequence SEQ ID NO: 134 (GI:88194185).

Useful sta099 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 134 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 134; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 134, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta099 proteins include variants of SEQ IDNO: 134. Preferred fragments of (b) comprise an epitope from SEQ ID NO:134. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 134 while retaining at leastone epitope of SEQ ID NO: 134. The first 30 N-terminal amino acids ofSEQ ID NO: 134 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta100

The ‘sta100’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta100 is SAOUHSC_(—)00391 and has amino acid sequenceSEQ ID NO: 135 (GI:88194186).

Useful sta100 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 135 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 135; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 135, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta100 proteins include variants of SEQ IDNO: 135. Preferred fragments of (b) comprise an epitope from SEQ ID NO:135. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 135 while retaining at leastone epitope of SEQ ID NO: 135. The first 30 N-terminal amino acids ofSEQ ID NO: 135 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta101

The ‘sta101’ antigen is annotated as ‘superantigen-like protein 7’. Inthe NCTC 8325 strain sta101 is SAOUHSC_(—)00392 and has amino acidsequence SEQ ID NO: 136 (GI:88194187). In the Newman strain it isnwmn_(—)0394 (GI:151220606).

Useful sta101 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 136 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 136; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 136, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta101 proteins include variants of SEQ IDNO: 136. Preferred fragments of (b) comprise an epitope from SEQ ID NO:136. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 136 while retaining at leastone epitope of SEQ ID NO: 136. The first 30 N-terminal amino acids ofSEQ ID NO: 136 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta102

The ‘sta102’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta102 is SAOUHSC_(—)00393 and has amino acid sequenceSEQ ID NO: 137 (GI:88194188).

Useful sta102 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 137 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 137; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 137, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta102 proteins include variants of SEQ IDNO: 137. Preferred fragments of (b) comprise an epitope from SEQ ID NO:137. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 137 while retaining at leastone epitope of SEQ ID NO: 137. The first 17 N-terminal amino acids ofSEQ ID NO: 137 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta103

The ‘sta103’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta103 is SAOUHSC_(—)00394 and has amino acid sequenceSEQ ID NO: 138 (GI:88194189).

Useful sta103 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 138 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 138; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 138, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta103 proteins include variants of SEQ IDNO: 138. Preferred fragments of (b) comprise an epitope from SEQ ID NO:138. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 138 while retaining at leastone epitope of SEQ ID NO: 138. The first 23 N-terminal amino acids ofSEQ ID NO: 138 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta104

The ‘sta104’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta104 is SAOUHSC_(—)00395 and has amino acid sequenceSEQ ID NO: 139 (GI:88194190).

Useful sta104 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 139 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 139; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 139, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta104 proteins include variants of SEQ IDNO: 139. Preferred fragments of (b) comprise an epitope from SEQ ID NO:139. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 139 while retaining at leastone epitope of SEQ ID NO: 139. Other fragments omit one or more proteindomains.

sta105

The ‘sta105’ antigen is annotated as ‘superantigen-like protein’. In theNCTC 8325 strain sta105 is SAOUHSC_(—)00399 and has amino acid sequenceSEQ ID NO: 140 (GI:88194194). In the Newman strain it is nwmn_(—)0400(GI:151220612).

Useful sta105 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 140 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 140; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 140, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta105 proteins include variants of SEQ IDNO: 140. Preferred fragments of (b) comprise an epitope from SEQ ID NO:140. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 140 while retaining at leastone epitope of SEQ ID NO: 140. The first 30 N-terminal amino acids ofSEQ ID NO: 140 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta106

The ‘sta106’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta106 is SAOUHSC_(—)01115 and has amino acid sequence SEQID NO: 141 (GI:88194861).

Useful sta106 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 141 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 141; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 141, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta106 proteins include variants of SEQ ID NO: 141.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 141.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 141 while retaining at least oneepitope of SEQ ID NO: 141. The first 16 N-terminal amino acids of SEQ IDNO: 141 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta107

The ‘sta107’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta107 is SAOUHSC_(—)00354 and has amino acid sequence SEQID NO: 177 (GI:88194153).

Useful sta107 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 177 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 177; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 177, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta107 proteins include variants of SEQ IDNO: 177. Preferred fragments of (b) comprise an epitope from SEQ ID NO:177. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 177 while retaining at leastone epitope of SEQ ID NO: 177. The first 35 N-terminal amino acids ofSEQ ID NO: 177 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta108

The ‘sta108’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta108 is SAOUHSC_(—)00717 and has amino acid sequence SEQID NO: 178 (GI:88194482).

Useful sta108 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 178 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 178; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 178, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta108 proteins include variants of SEQ ID NO: 178.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 178.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 178 while retaining at least oneepitope of SEQ ID NO: 178. The first 20 N-terminal amino acids of SEQ IDNO: 178 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta109

The ‘sta109’ antigen is annotated as ‘N-acetylmuramoyl-L-alanineamidase’. In the NCTC 8325 strain sta109 is SAOUHSC_(—)02979 and hasamino acid sequence SEQ ID NO: 179 (GI:88196599).

Useful sta109 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 179 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 179; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 179, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta109 proteins include variants ofSEQ ID NO: 179. Preferred fragments of (b) comprise an epitope from SEQID NO: 179. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 179 while retaining atleast one epitope of SEQ ID NO: 179. The first 27 N-terminal amino acidsof SEQ ID NO: 179 can usefully be omitted. Other fragments omit one ormore protein domains.

sta110

The ‘sta110’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta110 is SAOUHSC_(—)01039 and has amino acid sequence SEQID NO: 180 (GI:88194791).

Useful sta110 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 180 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 180; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 180, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These sta110 proteins include variants of SEQ IDNO: 180. Preferred fragments of (b) comprise an epitope from SEQ ID NO:180. Other preferred fragments lack one or more amino acids (e.g. 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/orone or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25or more) from the N-terminus of SEQ ID NO: 180 while retaining at leastone epitope of SEQ ID NO: 180. The first 19 N-terminal amino acids ofSEQ ID NO: 180 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta111

The ‘sta111’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta111 is SAOUHSC_(—)01005 and has amino acid sequence SEQID NO: 181 (GI:88194760).

Useful sta111 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 181 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 181; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 181, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta111 proteins include variants of SEQ ID NO: 181.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 181.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 181 while retaining at least oneepitope of SEQ ID NO: 181. The first 20 N-terminal amino acids of SEQ IDNO: 181 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta112

The ‘sta112’ antigen is annotated as a putative ‘ABC transporter,substrate-binding protein’. In the NCTC 8325 strain sta112 isSAOUHSC_(—)00634 and has amino acid sequence SEQ ID NO: 182(GI:88194402).

Useful sta112 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 182 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 182; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 182, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta112 proteins include variants ofSEQ ID NO: 182. Preferred fragments of (b) comprise an epitope from SEQID NO: 182. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 182 while retaining atleast one epitope of SEQ ID NO: 182. The first 17 N-terminal amino acidsof SEQ ID NO: 182 can usefully be omitted. Other fragments omit one ormore protein domains.

A Sta112 antigen can usefully be included in a composition incombination with a Sta073 [74].

sta113

The ‘sta113’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta113 is SAOUHSC_(—)00728 and has amino acid sequence SEQID NO: 183 (GI:88194493).

Useful sta113 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 183 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 183; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 183, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta113 proteins include variants ofSEQ ID NO: 183. Preferred fragments of (b) comprise an epitope from SEQID NO: 183. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 183 while retaining atleast one epitope of SEQ ID NO: 183. The first 173 N-terminal aminoacids of SEQ ID NO: 183 can usefully be omitted. Other fragments omitone or more protein domains.

sta114

The ‘sta114’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta114 is SAOUHSC_(—)00810 and has amino acid sequence SEQID NO: 184 (GI:88194570).

Useful sta114 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 184 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 184; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 184, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These sta114 proteins include variants of SEQ ID NO:184. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 184.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 184 while retaining at least oneepitope of SEQ ID NO: 184. Other fragments omit one or more proteindomains.

sta115

The ‘sta115’ antigen is annotated as ‘hypothetical protein’. In the NCTC8325 strain sta115 is SAOUHSC_(—)00817 and has amino acid sequence SEQID NO: 185 (GI:88194576).

Useful sta115 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 185 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 185; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 185, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These sta115 proteins include variants of SEQ ID NO:185. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 185.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 185 while retaining at least oneepitope of SEQ ID NO: 185. The first 18 N-terminal amino acids of SEQ IDNO: 185 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta116

The ‘sta116’ antigen is annotated as ‘formyl peptide receptor-like 1inhibitory protein’. In the NCTC 8325 strain sta116 is SAOUHSC_(—)01112and has amino acid sequence SEQ ID NO: 186 (GI:88194858).

Useful sta116 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 186 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 186; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 186, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta116 proteins include variants of SEQ ID NO: 186.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 186.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 186 while retaining at least oneepitope of SEQ ID NO: 186. The first 20 N-terminal amino acids of SEQ IDNO: 186 can usefully be omitted. Other fragments omit one or moreprotein domains.

sta117

The ‘sta117’ antigen is annotated as ‘truncated beta-hemolysin’. In theNCTC 8325 strain sta117 is SAOUHSC_(—)02240 and has amino acid sequenceSEQ ID NO: 187 (GI:88195913).

Useful sta117 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 187 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 187; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 187, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta117 proteins include variants ofSEQ ID NO: 187. Preferred fragments of (b) comprise an epitope from SEQID NO: 187. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 187 while retaining atleast one epitope of SEQ ID NO: 187. Other fragments omit one or moreprotein domains.

sta118

The ‘sta118’ antigen is annotated as ‘cell division protein FtsZ’. Inthe NCTC 8325 strain sta118 is SAOUHSC_(—)01150 and has amino acidsequence SEQ ID NO: 188 (GI:88194892).

Useful sta118 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 188 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 188; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 188, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These sta118 proteins include variants ofSEQ ID NO: 188. Preferred fragments of (b) comprise an epitope from SEQID NO: 188. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 188 while retaining atleast one epitope of SEQ ID NO: 188. Other fragments omit one or moreprotein domains.

sta119

The ‘sta119’ antigen is annotated as ‘thioredoxin’. In the NCTC 8325strain sta119 is SAOUHSC_(—)01100 and has amino acid sequence SEQ ID NO:200 (GI:88194846).

Useful sta119 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 200 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 200; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 200, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These sta119 proteins include variants of SEQ ID NO: 200.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 200.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 200 while retaining at least oneepitope of SEQ ID NO: 200. Other fragments omit one or more proteindomains.

sta120

The ‘sta120’ antigen is annotated as ‘alkyl hydroperoxide reductasesubunit C’. In the NCTC 8325 strain sta120 is SAOUHSC_(—)00365 and hasamino acid sequence SEQ ID NO: 201 (GI:88194163).

Useful sta120 antigens can elicit an antibody (e.g. when administered toa human) that recognises SEQ ID NO: 201 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 201; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 201, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These sta120 proteins include variants of SEQ ID NO:201. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 201.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 201 while retaining at least oneepitope of SEQ ID NO: 201. Other fragments omit one or more proteindomains.

NW_(—)6

The ‘NW_(—)6’ antigen is annotated as ‘secreted von Willebrandfactor-binding protein precursor’. In the Newman strain NW_(—)6 isNWMN_(—)0757 and has amino acid sequence SEQ ID NO: 142 (GI:151220969).

Useful NW_(—)6 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 142 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 142; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 142, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These NW_(—)6 proteins include variants ofSEQ ID NO: 142. Preferred fragments of (b) comprise an epitope from SEQID NO: 142. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 142 while retaining atleast one epitope of SEQ ID NO: 142. The first 13 N-terminal amino acidsof SEQ ID NO: 142 can usefully be omitted. Other fragments omit one ormore protein domains.

NW_(—)9

The ‘NW_(—)9’ antigen is annotated as ‘lipoprotein’. In the Newmanstrain NW_(—)9 is NWMN_(—)0958 and has amino acid sequence SEQ ID NO:143 (GI:151221170).

Useful NW_(—)9 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 143 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 143; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 143, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200 or more). These NW_(—)9 proteins include variants of SEQID NO: 143. Preferred fragments of (b) comprise an epitope from SEQ IDNO: 143. Other preferred fragments lack one or more amino acids (e.g. 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 143 while retaining atleast one epitope of SEQ ID NO: 143. The first 19 N-terminal amino acidsof SEQ ID NO: 143 can usefully be omitted. Other fragments omit one ormore protein domains.

NW_(—)10

The ‘NW_(—)10’ antigen is annotated as ‘fibrinogen binding-relatedprotein’. In the Newman strain NW_(—)10 is NWMN_(—)1066 and has aminoacid sequence SEQ ID NO: 144 (GI:151221278).

Useful NW_(—)10 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 144 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 144; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 144, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These NW_(—)10 proteins include variants of SEQ ID NO: 144.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 144.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 144 while retaining at least oneepitope of SEQ ID NO: 144. The first 20 N-terminal amino acids of SEQ IDNO: 144 can usefully be omitted. Other fragments omit one or moreprotein domains.

NW_(—)7

The ‘NW_(—)7’ antigen is annotated as ‘staphylococcal complementinhibitor SCIN’. In the Newman strain NW_(—)7 is NWMN_(—)1876 and hasamino acid sequence SEQ ID NO: 145 (GI:151222088).

Useful NW_(—)7 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 145 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 145; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 145, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These NW_(—)7 proteins include variants of SEQ ID NO: 145.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 145.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 145 while retaining at least oneepitope of SEQ ID NO: 145. The first 17 N-terminal amino acids of SEQ IDNO: 145 can usefully be omitted. Other fragments omit one or moreprotein domains.

NW_(—)8

The ‘NW_(—)8’ antigen is annotated as ‘chemotaxis-inhibiting proteinCHIPS’. In the Newman strain NW_(—)8 is NWMN_(—)1877 and has amino acidsequence SEQ ID NO: 146 (GI:151222089).

Useful NW_(—)8 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 146 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 146; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 146, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100or more). These NW_(—)8 proteins include variants of SEQ ID NO: 146.Preferred fragments of (b) comprise an epitope from SEQ ID NO: 146.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 146 while retaining at least oneepitope of SEQ ID NO: 146. The first 19 N-terminal amino acids of SEQ IDNO: 146 can usefully be omitted. Other fragments omit one or moreprotein domains.

NW_(—)2

The ‘NW_(—)2’ antigen is annotated as ‘enterotoxin type A precursor’. Inthe Newman strain NW_(—)2 is NWMN_(—)1883 and has amino acid sequenceSEQ ID NO: 147 (GI:151222095).

Useful NW_(—)2 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 147 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 147; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 147, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These NW_(—)2 proteins include variants ofSEQ ID NO: 147. Preferred fragments of (b) comprise an epitope from SEQID NO: 147. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 147 while retaining atleast one epitope of SEQ ID NO: 147. The first 16 N-terminal amino acidsof SEQ ID NO: 147 can usefully be omitted. Other fragments omit one ormore protein domains.

NW_(—)1

The ‘NW_(—)1’ antigen is annotated as ‘lipoprotein’. In the Newmanstrain NW_(—)1 is NWMN_(—)1924 and has amino acid sequence SEQ ID NO:148 (GI:151222136).

Useful NW_(—)1 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 148 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 148; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 148, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150 or more). These NW_(—)1 proteins include variants of SEQ ID NO:148. Preferred fragments of (b) comprise an epitope from SEQ ID NO: 148.Other preferred fragments lack one or more amino acids (e.g. 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminus and/or one ormore amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 ormore) from the N-terminus of SEQ ID NO: 148 while retaining at least oneepitope of SEQ ID NO: 148. The first 17 N-terminal amino acids of SEQ IDNO: 148 can usefully be omitted. Other fragments omit one or moreprotein domains.

NW_(—)5

The ‘NW_(—)5’ antigen is annotated as ‘cell wall surface anchor familyprotein’. In the Newman strain NW_(—)5 is NWMN_(—)2392 and has aminoacid sequence SEQ ID NO: 149 (GI:151222604).

Useful NW_(—)5 antigens can elicit an antibody (e.g. when administeredto a human) that recognises SEQ ID NO: 149 and/or may comprise an aminoacid sequence: (a) having 50% or more identity (e.g. 60%, 65%, 70%, 75%,80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% ormore) to SEQ ID NO: 149; and/or (b) comprising a fragment of at least‘n’ consecutive amino acids of SEQ ID NO: 149, wherein ‘n’ is 7 or more(e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90,100, 150, 200, 250 or more). These NW_(—)5 proteins include variants ofSEQ ID NO: 149. Preferred fragments of (b) comprise an epitope from SEQID NO: 149. Other preferred fragments lack one or more amino acids (e.g.1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from the C-terminusand/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15,20, 25 or more) from the N-terminus of SEQ ID NO: 149 while retaining atleast one epitope of SEQ ID NO: 149. The first 52 N-terminal amino acidsof SEQ ID NO: 149 can usefully be omitted. Other fragments omit one ormore protein domains.

Hybrid Polypeptides

Antigens used in the invention may be present in the composition asindividual separate polypeptides. Where more than one antigen is used,however, they do not have to be present as separate polypeptides.Instead, at least two (e.g. 2, 3, 4, 5, or more) antigens can beexpressed as a single polypeptide chain (a ‘hybrid’ polypeptide). Hybridpolypeptides offer two main advantages: first, a polypeptide that may beunstable or poorly expressed on its own can be assisted by adding asuitable hybrid partner that overcomes the problem; second, commercialmanufacture is simplified as only one expression and purification needbe employed in order to produce two polypeptides which are bothantigenically useful.

The hybrid polypeptide may comprise two or more polypeptide sequencesfrom the first antigen group. The hybrid polypeptide may comprise one ormore polypeptide sequences from the first antigen group and one or morepolypeptide sequences from the second antigen group. Moreover, thehybrid polypeptide may comprise two or more polypeptide sequences fromeach of the antigens listed above, or two or more variants of the sameantigen in the cases in which the sequence has partial variabilityacross strains.

Hybrids consisting of amino acid sequences from two, three, four, five,six, seven, eight, nine, or ten antigens are useful. In particular,hybrids consisting of amino acid sequences from two, three, four, orfive antigens are preferred, such as two or three antigens.

Different hybrid polypeptides may be mixed together in a singleformulation. Hybrids may be combined with non-hybrid antigens selectedfrom the first, second or third antigen groups. Within suchcombinations, an antigen may be present in more than one hybridpolypeptide and/or as a non-hybrid polypeptide. It is preferred,however, that an antigen is present either as a hybrid or as anon-hybrid, but not as both.

The hybrid polypeptides can also be combined with conjugates or non-S.aureus antigens as described above.

Hybrid polypeptides can be represented by the formulaNH₂-A-{-X-L-}_(n)-B—COOH, wherein: X is an amino acid sequence of a S.aureus antigen, as described above; L is an optional linker amino acidsequence; A is an optional N-terminal amino acid sequence; B is anoptional C-terminal amino acid sequence; n is an integer of 2 or more(e.g. 2, 3, 4, 5, 6, etc.). Usually n is 2 or 3.

If a —X— moiety has a leader peptide sequence in its wild-type form,this may be included or omitted in the hybrid protein. In someembodiments, the leader peptides will be deleted except for that of the—X— moiety located at the N-terminus of the hybrid protein i.e. theleader peptide of X₁ will be retained, but the leader peptides of X₂ . .. X_(n) will be omitted. This is equivalent to deleting all leaderpeptides and using the leader peptide of X₁ as moiety -A-.

For each n instances of {-X-L-}, linker amino acid sequence -L- may bepresent or absent. For instance, when n=2 the hybrid may beNH₂—X₁-L₁-X₂-L₂-COOH, NH₂—X₁—X₂—COOH, NH₂—X₁-L₁-X₂—COOH,NH₂—X₁—X₂-L₂-COOH, etc. Linker amino acid sequence(s)-L- will typicallybe short (e.g. 20 or fewer amino acids i.e. 20, 19, 18, 17, 16, 15, 14,13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples comprise shortpeptide sequences which facilitate cloning, poly-glycine linkers (i.e.comprising Gly_(n) where n=2, 3, 4, 5, 6, 7, 8, 9, 10 or more), andhistidine tags (i.e. His_(n) where n=3, 4, 5, 6, 7, 8, 9, 10 or more).Other suitable linker amino acid sequences will be apparent to thoseskilled in the art. A useful linker is GSGGGG (SEQ ID NO: 171) orGSGSGGGG (SEQ ID NO: 172), with the Gly-Ser dipeptide being formed froma BamHI restriction site (or two of them, to form the SEQ ID NO: 230tetrapeptide), thus aiding cloning and manipulation, and the (Gly)₄tetrapeptide (SEQ ID NO: 227) being a typical poly-glycine linker. Othersuitable linkers, particularly for use as the final L_(n) are ASGGGS(SEQ ID NO: 173 e.g. encoded by SEQ ID NO: 174) or a Leu-Glu dipeptide.

-A- is an optional N-terminal amino acid sequence. This will typicallybe short (e.g. 40 or fewer amino acids i.e. 40, 39, 38, 37, 36, 35, 34,33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16,15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples includeleader sequences to direct protein trafficking, or short peptidesequences which facilitate cloning or purification (e.g. histidine tagsi.e. His_(n) where n=3, 4, 5, 6, 7, 8, 9, 10 or more). Other suitableN-terminal amino acid sequences will be apparent to those skilled in theart. If X₁ lacks its own N-terminus methionine, -A- is preferably anoligopeptide (e.g. with 1, 2, 3, 4, 5, 6, 7 or 8 amino acids) whichprovides a N-terminus methionine e.g. Met-Ala-Ser, or a single Metresidue.

—B— is an optional C-terminal amino acid sequence. This will typicallybe short (e.g. 40 or fewer amino acids i.e. 39, 38, 37, 36, 35, 34, 33,32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15,14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1). Examples includesequences to direct protein trafficking, short peptide sequences whichfacilitate cloning or purification (e.g. comprising histidine tags i.e.His_(n) where n=3, 4, 5, 6, 7, 8, 9, 10 or more, such as SEQ ID NO:226), or sequences which enhance protein stability. Other suitableC-terminal amino acid sequences will be apparent to those skilled in theart.

One hybrid polypeptide of the invention may include both EsxA and EsxBantigens. These may be in either order, N- to C-terminus. SEQ ID NOs:151 (‘EsxAB’; encoded by SEQ ID NO: 169) and 152 (‘EsxBA’) are examplesof such hybrids, both having hexapeptide linkers ASGGGS (SEQ ID NO:173). Another ‘EsxAB’ hybrid comprises SEQ ID NO: 241, which may beprovided with a N-terminus methionine (e.g. SEQ ID NO: 250).

Another hybrid polypeptide of the invention may include both SdrD andSdrE antigens. These may be in either order, N- to C-terminus. SEQ IDNO: 168 (‘SdrED’) is an example of such a hybrid, having a hexapeptidelinker ASGGGS (SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include both ClfB andSdrD antigens. These may be in either order, N- to C-terminus. SEQ IDNO: 202 (‘ClfB-SdrD’) is an example of such a hybrid, having ahexapeptide linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 203 (‘SdrD-ClfB’)is another example of such a hybrid, having a hexapeptide linker ASGGGS(SEQ ID NO: 173). SEQ ID NO: 211 (‘ClfB-N-3-sdrD-N3’) is another exampleof such a hybrid, where the N3 fragments of ClfB and SdrD are joined byhexapeptide linker ASGGGS (SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include both IsdA andEsxA antigens. These may be in either order, N- to C-terminus. SEQ IDNO: 204 (‘IsdA-EsxA’) is an example of such a hybrid, having ahexapeptide linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 209(‘isdA40-184-esxA’) is another example of such a hybrid, in whichIsdA₄₀₋₁₈₄ is joined to EsxA via linker ASGGGS (SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include both IsdA andsta006 antigens. These may be in either order, N- to C-terminus. SEQ IDNO: 221 (‘isdA40-184-sta006’) is an example of such a hybrid, in whichIsdA₄₀₋₁₈₄ is joined to Sta006 via hexapeptide linker ASGGGS (SEQ ID NO:173).

Another hybrid polypeptide of the invention may include both Hla andsta006 antigens. These may be in either order, N- to C-terminus. SEQ IDNO: 222 (‘HlaH35L-sta006’) is an example of such a hybrid, in which aH35L mutant of Hla is joined to Sta006 via hexapeptide linker ASGGGS(SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include both Hla and Empantigens. These may be in either order, N- to C-terminus. SEQ ID NO: 205(‘HlaH35L-Emp’) is an example of such a hybrid, in which a H35L mutantHla is joined to Emp via linker ASGGGS (SEQ ID NO: 173). SEQ ID NO: 206(‘Hla27-76-Emp’) is another example of such a hybrid, in which a Hlafragment is joined to Emp via linker ASGGGS (SEQ ID NO: 173); SEQ ID NO:207 is a H35L mutant of SEQ ID NO: 206. SEQ ID NO: 208 (‘HlaPSGS-Emp’)is another example of such a hybrid, in which a Hla mutant is joined toEmp via linker ASGGGS (SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include IsdA and EsxAand EsxB antigens. These may be in any order, N- to C-terminus. SEQ IDNO: 210 (‘isdA40-184-esxAB’) is an example of such a triple hybrid, inwhich IsdA₄₀₋₁₈₄ is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173).The EsxAB already includes the same linker, so SEQ ID NO: 210 includestwo of these linkers. SEQ ID NO: 212 (‘IsdA-esxAB’) is another exampleof such a triple hybrid, in which IsdA is joined to EsxAB via linkerASGGGS (SEQ ID NO: 173).

Another hybrid polypeptide of the invention may include Hla and EsxA andEsxB antigens. These may be in any order, N- to C-terminus. SEQ ID NO:220 (‘HlaH35L-esxAB’) is an example of such a triple hybrid, in which aH35L mutant of Hla is joined to EsxAB via linker ASGGGS (SEQ ID NO:173). The EsxAB already includes the same linker, so SEQ ID NO: 220includes two of these linkers. Another example of a hybrid polypeptideincluding Hla and EsxA and EsxB antigens is SEQ ID NO: 237(‘HlaH35L-esxAB’ as used in the examples), in which a H35L mutant of Hlais joined to EsxA via linker APTARG (SEQ ID NO: 239) to replace itsN-terminus, then to EsxB via linker ASGGGS (SEQ ID NO: 173) to replaceits N-terminus. This hybrid can be provided with a suitable N-terminalsequence such as SEQ ID NO: 240.

Another hybrid polypeptide of the invention may include sta006 and EsxAand EsxB antigens. These may be in any order, N- to C-terminus. SEQ IDNO: 223 (‘sta006-esxAB’) is an example of such a triple hybrid, in whichsta006 is joined to EsxAB via linker ASGGGS (SEQ ID NO: 173). The EsxABalready includes the same linker, so SEQ ID NO: 223 includes two ofthese linkers. Another example of a hybrid polypeptide including sta006and EsxA and EsxB antigens is SEQ ID NO: 238 (‘sta006-esxAB’ as used inthe examples), in which a sta006 is joined to EsxA via linker APTARG(SEQ ID NO: 239) to replace its N-terminus, then to EsxB via linkerASGGGS (SEQ ID NO: 173) to replace its N-terminus. This hybrid can beprovided with a suitable N-terminal sequence such as SEQ ID NO: 240.

Usefully, these hybrid polypeptides can elicit an antibody (e.g. whenadministered to a human) that recognise each of the wild-typestaphylococcal proteins (e.g. as shown in the sequence listing)represented in the hybrid e.g. which recognise both wild-type EsxA andwild-type EsxB, or which recognise both wild-type SdrD and wild-typeSdrE, or which recognise both wild-type SdrD and wild-type ClfB, orwhich recognise both wild-type IsdA and wild-type EsxA, or whichrecognise both wild-type IsdA and wild-type sta006, or which recogniseboth wild-type Hla and wild-type sta006, or which recognise bothwild-type Hla and wild-type Emp, or which recognise wild-type IsdA andwild-type EsxA and wild-type EsxB, or which recognise wild-type Hla andwild-type EsxA and wild-type EsxB, or which recognise wild-type sta006and wild-type EsxA and wild-type EsxB.

Polypeptides Used with the Invention

Polypeptides used with the invention can take various forms (e.g.native, fusions, glycosylated, non-glycosylated, lipidated,non-lipidated, phosphorylated, non-phosphorylated, myristoylated,non-myristoylated, monomeric, multimeric, particulate, denatured, etc.).

Polypeptides used with the invention can be prepared by various means(e.g. recombinant expression, purification from cell culture, chemicalsynthesis, etc.). Recombinantly-expressed proteins are preferred,particularly for hybrid polypeptides.

Polypeptides used with the invention are preferably provided in purifiedor substantially purified form i.e. substantially free from otherpolypeptides (e.g. free from naturally-occurring polypeptides),particularly from other staphylococcal or host cell polypeptides, andare generally at least about 50% pure (by weight), and usually at leastabout 90% pure i.e. less than about 50%, and more preferably less thanabout 10% (e.g. 5%) of a composition is made up of other expressedpolypeptides. Thus the antigens in the compositions are separated fromthe whole organism with which the molecule is expressed.

Polypeptides used with the invention are preferably staphylococcalpolypeptides.

The term “polypeptide” refers to amino acid polymers of any length. Thepolymer may be linear or branched, it may comprise modified amino acids,and it may be interrupted by non-amino acids. The terms also encompassan amino acid polymer that has been modified naturally or byintervention; for example, disulfide bond formation, glycosylation,lipidation, acetylation, phosphorylation, or any other manipulation ormodification, such as conjugation with a labeling component. Alsoincluded are, for example, polypeptides containing one or more analogsof an amino acid (including, for example, unnatural amino acids, etc.),as well as other modifications known in the art. Polypeptides can occuras single chains or associated chains.

The invention provides polypeptides comprising a sequence —P-Q- or-Q-P—, wherein: —P— is an amino acid sequence as defined above and -Q-is not a sequence as defined above i.e. the invention provides fusionproteins. Where the N-terminus codon of —P— is not ATG, but this codonis not present at the N-terminus of a polypeptide, it will be translatedas the standard amino acid for that codon rather than as a Met. Wherethis codon is at the N-terminus of a polypeptide, however, it will betranslated as Met. Examples of -Q- moieties include, but are not limitedto, histidine tags (i.e. His_(n) where n=3, 4, 5, 6, 7, 8, 9, 10 ormore), maltose-binding protein, or glutathione-S-transferase (GST).

The invention also provides a process for producing a polypeptide of theinvention, comprising the step of culturing a host cell transformed withnucleic acid of the invention under conditions which induce polypeptideexpression.

Although expression of the polypeptides of the invention may take placein a Staphylococcus, the invention will usually use a heterologous hostfor expression (recombinant expression). The heterologous host may beprokaryotic (e.g. a bacterium) or eukaryotic. It may be E. coli, butother suitable hosts include Bacillus subtilis, Vibrio cholerae,Salmonella typhi, Salmonella typhimurium, Neisseria lactamica, Neisseriacinerea, Mycobacteria (e.g. M. tuberculosis), yeasts, etc. Compared tothe wild-type S. aureus genes encoding polypeptides of the invention, itis helpful to change codons to optimise expression efficiency in suchhosts without affecting the encoded amino acids.

The invention provides a process for producing a polypeptide of theinvention, comprising the step of synthesising at least part of thepolypeptide by chemical means.

Nucleic Acids

The invention also provides nucleic acid encoding polypeptides andhybrid polypeptides of the invention. It also provides nucleic acidcomprising a nucleotide sequence that encodes one or more polypeptidesor hybrid polypeptides of the invention.

The invention also provides nucleic acid comprising nucleotide sequenceshaving sequence identity to such nucleotide sequences. Identity betweensequences is preferably determined by the Smith-Waterman homology searchalgorithm as described above. Such nucleic acids include those usingalternative codons to encode the same amino acid.

The invention also provides nucleic acid which can hybridize to thesenucleic acids. Hybridization reactions can be performed under conditionsof different “stringency”. Conditions that increase stringency of ahybridization reaction of widely known and published in the art (e.g.page 7.52 of reference 276). Examples of relevant conditions include (inorder of increasing stringency): incubation temperatures of 25° C., 37°C., 50° C., 55° C. and 68° C.; buffer concentrations of 10×SSC, 6×SSC,1×SSC, 0.1×SSC (where SSC is 0.15 M NaCl and 15 mM citrate buffer) andtheir equivalents using other buffer systems; formamide concentrationsof 0%, 25%, 50%, and 75%; incubation times from 5 minutes to 24 hours;1, 2, or more washing steps; wash incubation times of 1, 2, or 15minutes; and wash solutions of 6×SSC, 1×SSC, 0.1×SSC, or de-ionizedwater. Hybridization techniques and their optimization are well known inthe art (e.g. see refs 75, 76, 276, 278, etc.].

In some embodiments, nucleic acid of the invention hybridizes to atarget under low stringency conditions; in other embodiments ithybridizes under intermediate stringency conditions; in preferredembodiments, it hybridizes under high stringency conditions. Anexemplary set of low stringency hybridization conditions is 50° C. and10×SSC. An exemplary set of intermediate stringency hybridizationconditions is 55° C. and 1×SSC. An exemplary set of high stringencyhybridization conditions is 68° C. and 0.1×SSC.

The invention includes nucleic acid comprising sequences complementaryto these sequences (e.g. for antisense or probing, or for use asprimers).

Nucleic acids of the invention can be used in hybridisation reactions(e.g. Northern or Southern blots, or in nucleic acid microarrays or‘gene chips’) and amplification reactions (e.g. PCR, SDA, SSSR, LCR,TMA, NASBA, etc.) and other nucleic acid techniques.

Nucleic acid according to the invention can take various forms (e.g.single-stranded, double-stranded, vectors, primers, probes, labelledetc.). Nucleic acids of the invention may be circular or branched, butwill generally be linear. Unless otherwise specified or required, anyembodiment of the invention that utilizes a nucleic acid may utilizeboth the double-stranded form and each of two complementarysingle-stranded forms which make up the double-stranded form. Primersand probes are generally single-stranded, as are antisense nucleicacids.

Nucleic acids of the invention are preferably provided in purified orsubstantially purified form i.e. substantially free from other nucleicacids (e.g. free from naturally-occurring nucleic acids), particularlyfrom other staphylococcal or host cell nucleic acids, generally being atleast about 50% pure (by weight), and usually at least about 90% pure.Nucleic acids of the invention are preferably staphylococcal nucleicacids.

Nucleic acids of the invention may be prepared in many ways e.g. bychemical synthesis (e.g. phosphoramidite synthesis of DNA) in whole orin part, by digesting longer nucleic acids using nucleases (e.g.restriction enzymes), by joining shorter nucleic acids or nucleotides(e.g. using ligases or polymerases), from genomic or cDNA libraries,etc.

Nucleic acid of the invention may be attached to a solid support (e.g. abead, plate, filter, film, slide, microarray support, resin, etc.).Nucleic acid of the invention may be labelled e.g. with a radioactive orfluorescent label, or a biotin label. This is particularly useful wherethe nucleic acid is to be used in detection techniques e.g. where thenucleic acid is a primer or as a probe.

The term “nucleic acid” includes in general means a polymeric form ofnucleotides of any length, which contain deoxyribonucleotides,ribonucleotides, and/or their analogs. It includes DNA, RNA, DNA/RNAhybrids. It also includes DNA or RNA analogs, such as those containingmodified backbones (e.g. peptide nucleic acids (PNAs) orphosphorothioates) or modified bases. Thus the invention includes mRNA,tRNA, rRNA, ribozymes, DNA, cDNA, recombinant nucleic acids, branchednucleic acids, plasmids, vectors, probes, primers, etc. Where nucleicacid of the invention takes the form of RNA, it may or may not have a 5′cap.

Nucleic acids of the invention may be part of a vector i.e. part of anucleic acid construct designed for transduction/transfection of one ormore cell types. Vectors may be, for example, “cloning vectors” whichare designed for isolation, propagation and replication of insertednucleotides, “expression vectors” which are designed for expression of anucleotide sequence in a host cell, “viral vectors” which is designed toresult in the production of a recombinant virus or virus-like particle,or “shuttle vectors”, which comprise the attributes of more than onetype of vector. Preferred vectors are plasmids. A “host cell” includesan individual cell or cell culture which can be or has been a recipientof exogenous nucleic acid. Host cells include progeny of a single hostcell, and the progeny may not necessarily be completely identical (inmorphology or in total DNA complement) to the original parent cell dueto natural, accidental, or deliberate mutation and/or change. Host cellsinclude cells transfected or infected in vivo or in vitro with nucleicacid of the invention.

Where a nucleic acid is DNA, it will be appreciated that “U” in a RNAsequence will be replaced by “T” in the DNA. Similarly, where a nucleicacid is RNA, it will be appreciated that “T” in a DNA sequence will bereplaced by “U” in the RNA.

The term “complement” or “complementary” when used in relation tonucleic acids refers to Watson-Crick base pairing. Thus the complementof C is G, the complement of G is C, the complement of A is T (or U),and the complement of T (or U) is A. It is also possible to use basessuch as I (the purine inosine) e.g. to complement pyrimidines (C or T).

Nucleic acids of the invention can be used, for example: to producepolypeptides; as hybridization probes for the detection of nucleic acidin biological samples; to generate additional copies of the nucleicacids; to generate ribozymes or antisense oligonucleotides; assingle-stranded DNA primers or probes; or as triple-strand formingoligonucleotides.

The invention provides a process for producing nucleic acid of theinvention, wherein the nucleic acid is synthesised in part or in wholeusing chemical means.

The invention provides vectors comprising nucleotide sequences of theinvention (e.g. cloning or expression vectors) and host cellstransformed with such vectors.

Nucleic acid amplification according to the invention may bequantitative and/or real-time.

For certain embodiments of the invention, nucleic acids are preferablyat least 7 nucleotides in length (e.g. 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70, 75, 80, 90, 100, 110,120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300nucleotides or longer).

For certain embodiments of the invention, nucleic acids are preferablyat most 500 nucleotides in length (e.g. 450, 400, 350, 300, 250, 200,150, 140, 130, 120, 110, 100, 90, 80, 75, 70, 65, 60, 55, 50, 45, 40,39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22,21, 20, 19, 18, 17, 16, 15 nucleotides or shorter).

Primers and probes of the invention, and other nucleic acids used forhybridization, are preferably between 10 and 30 nucleotides in length(e.g. 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, or 30 nucleotides).

Strains and Variants

Antigens are defined above by reference to existing nomenclature (e.g.“ClfA”), to “sta” numbers or to “NW_” numbers. Table 1 herein relatesthese three naming/numbering systems to existing SAOUHSC numberingand/or NWMN numbering. SAOUHSC numbering refers to the genome of S.aureus strain NCTC 8325 (sequenced by Oklahoma University HealthSciences Center and disclosed in GenBank as CP000253.1; GI:87201381),and individual SAOUHSC numbers are given as “locus_tag” entries in thegenome sequence's “features” section. Similarly, NWMN numbering refersto the genome of S. aureus strain Newman (isolated in 1952 from a humaninfection, and having robust virulence phenotype) disclosed in GenBankas AP009351.1 (GI:150373012) and individual NWMN numbers are given as“locus_tag” entries in the genome sequence's “features” section.Functional annotations for each antigen are also given in the databases.

Table 1 also includes the GI number for each antigen of the invention.Thus an exemplary amino acid and nucleotide sequence for any of theseantigens can easily be found in public sequence databases from the NCTC8325 and/or Newman strain, but the invention is not limited to sequencesfrom the NCTC 8325 and Newman strains. Genome sequences of several otherstrains of S. aureus are available, including those of MRSA strains N315and Mu50 [77], MW2, N315, COL, MRSA252, MSSA476, RF122, USA300 (veryvirulent), JH1 and JH9. Standard search and alignment techniques can beused to identify in any of these (or other) further genome sequences thehomolog of any particular sequence from the Newman or NCTC 8325 strain.Moreover, the available sequences from the Newman and NCTC 8325 strainscan be used to design primers for amplification of homologous sequencesfrom other strains. Thus the invention is not limited to these twostrains, but rather encompasses such variants and homologs from otherstrains of S. aureus, as well as non-natural variants. In general,suitable variants of a particular SEQ ID NO include its allelicvariants, its polymorphic forms, its homologs, its orthologs, itsparalogs, its mutants, etc.

Thus, for instance, polypeptides used with the invention may, comparedto the SEQ ID NO herein, include one or more (e.g. 1, 2, 3, 4, 5, 6, 7,8, 9, etc.) amino acid substitutions, such as conservative substitutions(i.e. substitutions of one amino acid with another which has a relatedside chain). Genetically-encoded amino acids are generally divided intofour families: (1) acidic i.e. aspartate, glutamate; (2) basic i.e.lysine, arginine, histidine; (3) non-polar i.e. alanine, valine,leucine, isoleucine, proline, phenylalanine, methionine, tryptophan; and(4) uncharged polar i.e. glycine, asparagine, glutamine, cysteine,serine, threonine, tyrosine. Phenylalanine, tryptophan, and tyrosine aresometimes classified jointly as aromatic amino acids. In general,substitution of single amino acids within these families does not have amajor effect on the biological activity. The polypeptides may alsoinclude one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.) single aminoacid deletions relative to the SEQ ID NO sequences. The polypeptides mayalso include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.)insertions (e.g. each of 1, 2, 3, 4 or 5 amino acids) relative to theSEQ ID NO sequences.

Similarly, a polypeptide used with the invention may comprise an aminoacid sequence that:

is identical (i.e. 100% identical) to a sequence disclosed in thesequence listing;shares sequence identity (e.g. 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99%, 99.5% or more) with a sequence disclosed in thesequence listing;has 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 (or more) single amino acidalterations (deletions, insertions, substitutions), which may be atseparate locations or may be contiguous, as compared to the sequences of(a) or (b); andwhen aligned with a particular sequence from the sequence listing usinga pairwise alignment algorithm, each moving window of x amino acids fromN-terminus to C-terminus (such that for an alignment that extends to pamino acids, where p>x, there are p−x+1 such windows) has at least xyidentical aligned amino acids, where: x is selected from 20, 25, 30, 35,40, 45, 50, 60, 70, 80, 90, 100, 150, 200; y is selected from 0.50,0.60, 0.70, 0.75, 0.80, 0.85, 0.90, 0.91, 0.92, 0.93, 0.94, 0.95, 0.96,0.97, 0.98, 0.99; and if x·y is not an integer then it is rounded up tothe nearest integer. The preferred pairwise alignment algorithm is theNeedleman-Wunsch global alignment algorithm [78], using defaultparameters (e.g. with Gap opening penalty=10.0, and with Gap extensionpenalty=0.5, using the EBLOSUM62 scoring matrix). This algorithm isconveniently implemented in the needle tool in the EMBOSS package [79].

Where hybrid polypeptides are used, the individual antigens within thehybrid (i.e. individual —X— moieties) may be from one or more strains.Where n=2, for instance, X₂ may be from the same strain as X₁ or from adifferent strain. Where n=3, the strains might be (i) X₁=X₂=X₃ (ii)X₁=X₂≠X₃ (iii) X₁≠X₂=X₃ (iv) X₁≠X₂≠₃ or (v) X₁=X₃≠X₂, etc.

Within group (c), deletions or substitutions may be at the N-terminusand/or C-terminus, or may be between the two termini. Thus a truncationis an example of a deletion. Truncations may involve deletion of up to40 (or more) amino acids at the N-terminus and/or C-terminus. N-terminustruncation can remove leader peptides e.g. to facilitate recombinantexpression in a heterologous host. C-terminus truncation can removeanchor sequences e.g. to facilitate recombinant expression in aheterologous host.

In general, when an antigen comprises a sequence that is not identicalto a complete S. aureus sequence from the sequence listing (e.g. when itcomprises a sequence listing with <100% sequence identity thereto, orwhen it comprises a fragment thereof) it is preferred in each individualinstance that the antigen can elicit an antibody which recognises therespective complete S. aureus sequence.

Mutant Bacteria

The invention also provides a S. aureus bacterium in which one or moreof the antigens from the various antigen groups of the inventionhas/have been knocked out. Techniques for producing knockout bacteriaare well known, and knockout S. aureus strains have been reported. Aknockout mutation may be situated in the coding region of the gene ormay lie within its transcriptional control regions (e.g. within itspromoter). A knockout mutation will reduce the level of mRNA encodingthe antigen to <1% of that produced by the wild-type bacterium,preferably <0.5%, more preferably <0.1%, and most preferably to 0%.

The invention also provides a S. aureus in which one or more of theantigens from the various antigen groups of the invention has a mutationwhich inhibits its activity. The gene encoding the antigen will have amutation that changes the encoded amino acid sequence. Mutation mayinvolve deletion, substitution, and/or insertion, any of which may beinvolve one or more amino acids.

The invention also provides a bacterium, such as a S. aureus bacterium,which hyper-expresses an antigen of the invention.

The invention also provides a bacterium, such as a S. aureus bacterium,that constitutively expresses an antigen of the invention. The inventionalso provides a meningococcus comprising a gene encoding an antigen ofthe invention, wherein the gene is under the control of an induciblepromoter.

Immunogenic Compositions and Medicaments

Immunogenic compositions of the invention may be useful as vaccines.Vaccines according to the invention may either be prophylactic (i.e. toprevent infection) or therapeutic (i.e. to treat infection), but willtypically be prophylactic.

Compositions may thus be pharmaceutically acceptable. They will usuallyinclude components in addition to the antigens e.g. they typicallyinclude one or more pharmaceutical carrier(s) and/or excipient(s). Athorough discussion of such components is available in reference 273.

Compositions will generally be administered to a mammal in aqueous form.Prior to administration, however, the composition may have been in anon-aqueous form. For instance, although some vaccines are manufacturedin aqueous form, then filled and distributed and administered also inaqueous form, other vaccines are lyophilised during manufacture and arereconstituted into an aqueous form at the time of use. Thus acomposition of the invention may be dried, such as a lyophilisedformulation.

The composition may include preservatives such as thiomersal or2-phenoxyethanol. It is preferred, however, that the vaccine should besubstantially free from (i.e. less than 5 μg/ml) mercurial material e.g.thiomersal-free. Vaccines containing no mercury are more preferred.Preservative-free vaccines are particularly preferred.

To improve thermal stability, a composition may include a temperatureprotective agent. Further details of such agents are provided below.

To control tonicity, it is preferred to include a physiological salt,such as a sodium salt. Sodium chloride (NaCl) is preferred, which may bepresent at between 1 and 20 mg/ml e.g. about 10±2 mg/ml NaCl. Othersalts that may be present include potassium chloride, potassiumdihydrogen phosphate, disodium phosphate dehydrate, magnesium chloride,calcium chloride, etc.

Compositions will generally have an osmolality of between 200 mOsm/kgand 400 mOsm/kg, preferably between 240-360 mOsm/kg, and will morepreferably fall within the range of 290-310 mOsm/kg.

Compositions may include one or more buffers. Typical buffers include: aphosphate buffer; a Tris buffer; a borate buffer; a succinate buffer; ahistidine buffer (particularly with an aluminum hydroxide adjuvant); ora citrate buffer. Buffers will typically be included in the 5-20 mMrange.

The pH of a composition will generally be between 5.0 and 8.1, and moretypically between 6.0 and 8.0 e.g. 6.5 and 7.5, or between 7.0 and 7.8.

The composition is preferably sterile. The composition is preferablynon-pyrogenic e.g. containing <1 EU (endotoxin unit, a standard measure)per dose, and preferably <0.1 EU per dose. The composition is preferablygluten free.

The composition may include material for a single immunisation, or mayinclude material for multiple immunisations (i.e. a ‘multidose’ kit).The inclusion of a preservative is preferred in multidose arrangements.As an alternative (or in addition) to including a preservative inmultidose compositions, the compositions may be contained in a containerhaving an aseptic adaptor for removal of material.

Human vaccines are typically administered in a dosage volume of about0.5 ml, although a half dose (i.e. about 0.25 ml) may be administered tochildren.

Immunogenic compositions of the invention may also comprise one or moreimmunoregulatory agents. Preferably, one or more of the immunoregulatoryagents include one or more adjuvants. The adjuvants may include a TH1adjuvant and/or a TH2 adjuvant, further discussed below.

Thus the invention provides an immunogenic composition comprising acombination of:

-   -   (1) one or more antigen(s) selected from the first, second,        third and fourth antigen groups (as defined above); and    -   (2) an adjuvant, such as an aluminium hydroxide adjuvant (for        example, one or more antigens may be adsorbed to aluminium        hydroxide).

For instance, the invention provides an immunogenic compositioncomprising a combination of a sta006 antigen and an adjuvant, such as analuminium hydroxide adjuvant. Similarly, the invention provides animmunogenic composition comprising a combination of a sta011 antigen andan adjuvant, such as an aluminium hydroxide adjuvant. These compositionsare ideally buffered e.g. with a histidine buffer.

Adjuvants which may be used in compositions of the invention include,but are not limited to:

A. Mineral-Containing Compositions

Mineral containing compositions suitable for use as adjuvants in theinvention include mineral salts, such as aluminium salts and calciumsalts (or mixtures thereof). Calcium salts include calcium phosphate(e.g. the “CAP” particles disclosed in ref. 80). Aluminum salts includehydroxides, phosphates, sulfates, etc., with the salts taking anysuitable form (e.g. gel, crystalline, amorphous, etc.). Adsorption tothese salts is preferred (e.g. all antigens may be adsorbed). Themineral containing compositions may also be formulated as a particle ofmetal salt [81].

The adjuvants known as aluminum hydroxide and aluminum phosphate may beused. These names are conventional, but are used for convenience only,as neither is a precise description of the actual chemical compoundwhich is present (e.g. see chapter 9 of reference 82)). The inventioncan use any of the “hydroxide” or “phosphate” adjuvants that are ingeneral use as adjuvants. The adjuvants known as “aluminium hydroxide”are typically aluminium oxyhydroxide salts, which are usually at leastpartially crystalline. The adjuvants known as “aluminium phosphate” aretypically aluminium hydroxyphosphates, often also containing a smallamount of sulfate (i.e. aluminium hydroxyphosphate sulfate). They may beobtained by precipitation, and the reaction conditions andconcentrations during precipitation influence the degree of substitutionof phosphate for hydroxyl in the salt.

A fibrous morphology (e.g. as seen in transmission electron micrographs)is typical for aluminium hydroxide adjuvants. The pI of aluminiumhydroxide adjuvants is typically about 11 i.e. the adjuvant itself has apositive surface charge at physiological pH. Adsorptive capacities ofbetween 1.8-2.6 mg protein per mg Al⁺⁺⁺ at pH 7.4 have been reported foraluminium hydroxide adjuvants.

Aluminium phosphate adjuvants generally have a PO₄/Al molar ratiobetween 0.3 and 1.2, preferably between 0.8 and 1.2, and more preferably0.95+0.1. The aluminium phosphate will generally be amorphous,particularly for hydroxyphosphate salts. A typical adjuvant is amorphousaluminium hydroxyphosphate with PO₄/Al molar ratio between 0.84 and0.92, included at 0.6 mg Al³⁺/ml. The aluminium phosphate will generallybe particulate (e.g. plate-like morphology as seen in transmissionelectron micrographs). Typical diameters of the particles are in therange 0.5-20 μm (e.g. about 5-10 μm) after any antigen adsorption.Adsorptive capacities of between 0.7-1.5 mg protein per mg Al⁺⁺⁺ at pH7.4 have been reported for aluminium phosphate adjuvants.

The point of zero charge (PZC) of aluminium phosphate is inverselyrelated to the degree of substitution of phosphate for hydroxyl, andthis degree of substitution can vary depending on reaction conditionsand concentration of reactants used for preparing the salt byprecipitation. PZC is also altered by changing the concentration of freephosphate ions in solution (more phosphate=more acidic PZC) or by addinga buffer such as a histidine buffer (makes PZC more basic). Aluminiumphosphates used according to the invention will generally have a PZC ofbetween 4.0 and 7.0, more preferably between 5.0 and 6.5 e.g. about 5.7.

As shown below, adsorption of S. aureus protein antigens (except IsdA,Sta019 and Sta073) to an aluminium hydroxide adjuvant is advantageous,particularly in a multi-protein combination (in which all antigens maybe adsorbed). A histidine buffer can usefully be included in suchadjuvanted compositions.

Suspensions of aluminium salts used to prepare compositions of theinvention may contain a buffer (e.g. a phosphate or a histidine or aTris buffer), but this is not always necessary. The suspensions arepreferably sterile and pyrogen-free. A suspension may include freeaqueous phosphate ions e.g. present at a concentration between 1.0 and20 mM, preferably between 5 and 15 mM, and more preferably about 10 mM.The suspensions may also comprise sodium chloride.

The invention can use a mixture of both an aluminium hydroxide and analuminium phosphate. In this case there may be more aluminium phosphatethan hydroxide e.g. a weight ratio of at least 2:1 e.g. ≧5:1, ≧6:1,≧7:1, ≧8:1, ≧9:1, etc.

The concentration of Al⁺⁺⁺ in a composition for administration to apatient is preferably less than 10 mg/ml e.g. ≦5 mg/ml, ≦4 mg/ml, ≦3mg/ml, ≦2 mg/ml, ≦1 mg/ml, etc. A preferred range is between 0.3 and 1mg/ml. A maximum of 0.85 mg/dose is preferred.

B. Oil Emulsions

Oil emulsion compositions suitable for use as adjuvants in the inventioninclude squalene-water emulsions, such as MF59 [Chapter 10 of ref. 82;see also ref. 83] (5% Squalene, 0.5% Tween 80, and 0.5% Span 85,formulated into submicron particles using a microfluidizer). CompleteFreund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA) may alsobe used.

Various oil-in-water emulsion adjuvants are known, and they typicallyinclude at least one oil and at least one surfactant, with the oil(s)and surfactant(s) being biodegradable (metabolisable) and biocompatible.The oil droplets in the emulsion are generally less than 5 μm indiameter, and ideally have a sub-micron diameter, with these small sizesbeing achieved with a microfluidiser to provide stable emulsions.Droplets with a size less than 220 nm are preferred as they can besubjected to filter sterilization.

The emulsion can comprise oils such as those from an animal (such asfish) or vegetable source. Sources for vegetable oils include nuts,seeds and grains. Peanut oil, soybean oil, coconut oil, and olive oil,the most commonly available, exemplify the nut oils. Jojoba oil can beused e.g. obtained from the jojoba bean. Seed oils include saffloweroil, cottonseed oil, sunflower seed oil, sesame seed oil and the like.In the grain group, corn oil is the most readily available, but the oilof other cereal grains such as wheat, oats, rye, rice, teff, triticaleand the like may also be used. 6-10 carbon fatty acid esters of glyceroland 1,2-propanediol, while not occurring naturally in seed oils, may beprepared by hydrolysis, separation and esterification of the appropriatematerials starting from the nut and seed oils. Fats and oils frommammalian milk are metabolizable and may therefore be used in thepractice of this invention. The procedures for separation, purification,saponification and other means necessary for obtaining pure oils fromanimal sources are well known in the art. Most fish containmetabolizable oils which may be readily recovered. For example, codliver oil, shark liver oils, and whale oil such as spermaceti exemplifyseveral of the fish oils which may be used herein. A number of branchedchain oils are synthesized biochemically in 5-carbon isoprene units andare generally referred to as terpenoids. Shark liver oil contains abranched, unsaturated terpenoids known as squalene,2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene, which isparticularly preferred herein. Squalane, the saturated analog tosqualene, is also a preferred oil. Fish oils, including squalene andsqualane, are readily available from commercial sources or may beobtained by methods known in the art. Other preferred oils are thetocopherols (see below). Mixtures of oils can be used.

Surfactants can be classified by their ‘HLB’ (hydrophile/lipophilebalance). Preferred surfactants of the invention have a HLB of at least10, preferably at least 15, and more preferably at least 16. Theinvention can be used with surfactants including, but not limited to:the polyoxyethylene sorbitan esters surfactants (commonly referred to asthe Tweens), especially polysorbate 20 and polysorbate 80; copolymers ofethylene oxide (EO), propylene oxide (PO), and/or butylene oxide (BO),sold under the DOWFAX™ tradename, such as linear EO/PO block copolymers;octoxynols, which can vary in the number of repeating ethoxy(oxy-1,2-ethanediyl) groups, with octoxynol-9 (Triton X-100, ort-octylphenoxypolyethoxyethanol) being of particular interest;(octylphenoxy)polyethoxyethanol (IGEPAL CA-630/NP-40); phospholipidssuch as phosphatidylcholine (lecithin); nonylphenol ethoxylates, such asthe Tergitol™ NP series; polyoxyethylene fatty ethers derived fromlauryl, cetyl, stearyl and oleyl alcohols (known as Brij surfactants),such as triethyleneglycol monolauryl ether (Brij 30); and sorbitanesters (commonly known as the SPANs), such as sorbitan trioleate (Span85) and sorbitan monolaurate. Non-ionic surfactants are preferred.Preferred surfactants for including in the emulsion are Tween 80(polyoxyethylene sorbitan monooleate), Span 85 (sorbitan trioleate),lecithin and Triton X-100.

Mixtures of surfactants can be used e.g. Tween 80/Span 85 mixtures. Acombination of a polyoxyethylene sorbitan ester such as polyoxyethylenesorbitan monooleate (Tween 80) and an octoxynol such ast-octylphenoxypolyethoxyethanol (Triton X-100) is also suitable. Anotheruseful combination comprises laureth 9 plus a polyoxyethylene sorbitanester and/or an octoxynol.

Preferred amounts of surfactants (% by weight) are: polyoxyethylenesorbitan esters (such as Tween 80) 0.01 to 1%, in particular about 0.1%;octyl- or nonylphenoxy polyoxyethanols (such as Triton X-100, or otherdetergents in the Triton series) 0.001 to 0.1%, in particular 0.005 to0.02%; polyoxyethylene ethers (such as laureth 9) 0.1 to 20%, preferably0.1 to 10% and in particular 0.1 to 1% or about 0.5%.

Preferred emulsion adjuvants have an average droplets size of ≦1 μm e.g.≦750 nm, ≦500 nm, ≦400 nm, ≦300 nm, ≦250 nm, ≦220 nm, ≦200 nm, orsmaller. These droplet sizes can conveniently be achieved by techniquessuch as microfluidisation.

Specific oil-in-water emulsion adjuvants useful with the inventioninclude, but are not limited to:

-   -   A submicron emulsion of squalene, Tween 80, and Span 85. The        composition of the emulsion by volume can be about 5% squalene,        about 0.5% polysorbate 80 and about 0.5% Span 85. In weight        terms, these ratios become 4.3% squalene, 0.5% polysorbate 80        and 0.48% Span 85. This adjuvant is known as ‘MF59’ [84-86], as        described in more detail in Chapter 10 of ref 87 and chapter 12        of ref 88. The MF59 emulsion advantageously includes citrate        ions e.g. 10 mM sodium citrate buffer.    -   An emulsion of squalene, a tocopherol, and polysorbate 80 (Tween        80). The emulsion may include phosphate buffered saline. It may        also include Span 85 (e.g. at 1%) and/or lecithin. These        emulsions may have from 2 to 10% squalene, from 2 to 10%        tocopherol and from 0.3 to 3% Tween 80, and the weight ratio of        squalene:tocopherol is preferably ≦1 as this provides a more        stable emulsion. Squalene and Tween 80 may be present volume        ratio of about 5:2 or at a weight ratio of about 11:5. One such        emulsion can be made by dissolving Tween 80 in PBS to give a 2%        solution, then mixing 90 ml of this solution with a mixture of        (5 g of DL-α-tocopherol and 5 ml squalene), then microfluidising        the mixture. The resulting emulsion may have submicron oil        droplets e.g. with an average diameter of between 100 and 250        nm, preferably about 180 nm. The emulsion may also include a        3-de-O-acylated monophosphoryl lipid A (3d-MPL). Another useful        emulsion of this type may comprise, per human dose, 0.5-10 mg        squalene, 0.5-11 mg tocopherol, and 0.1-4 mg polysorbate 80        [89].    -   An emulsion of squalene, a tocopherol, and a Triton detergent        (e.g. Triton X-100). The emulsion may also include a 3d-MPL (see        below). The emulsion may contain a phosphate buffer.    -   An emulsion comprising a polysorbate (e.g. polysorbate 80), a        Triton detergent (e.g. Triton X-100) and a tocopherol (e.g. an        α-tocopherol succinate). The emulsion may include these three        components at a mass ratio of about 75:11:10 (e.g. 750 μg/ml        polysorbate 80, 110 m/ml Triton X-100 and 100 μg/ml α-tocopherol        succinate), and these concentrations should include any        contribution of these components from antigens. The emulsion may        also include squalene. The emulsion may also include a 3d-MPL        (see below). The aqueous phase may contain a phosphate buffer.    -   An emulsion of squalane, polysorbate 80 and poloxamer 401        (“Pluronic™ L121”). The emulsion can be formulated in phosphate        buffered saline, pH 7.4. This emulsion is a useful delivery        vehicle for muramyl dipeptides, and has been used with        threonyl-MDP in the “SAF-1” adjuvant [90] (0.05-1% Thr-MDP, 5%        squalane, 2.5% Pluronic L121 and 0.2% polysorbate 80). It can        also be used without the Thr-MDP, as in the “AF” adjuvant [91]        (5% squalane, 1.25% Pluronic L121 and 0.2% polysorbate 80).        Microfluidisation is preferred.    -   An emulsion comprising squalene, an aqueous solvent, a        polyoxyethylene alkyl ether hydrophilic nonionic surfactant        (e.g. polyoxyethylene (12) cetostearyl ether) and a hydrophobic        nonionic surfactant (e.g. a sorbitan ester or mannide ester,        such as sorbitan monoleate or ‘Span 80’). The emulsion is        preferably thermoreversible and/or has at least 90% of the oil        droplets (by volume) with a size less than 200 nm [92]. The        emulsion may also include one or more of alditol; a        cryoprotective agent (e.g. a sugar, such as dodecylmaltoside        and/or sucrose); and/or an alkylpolyglycoside. The emulsion may        include a TLR4 agonist [93]. Such emulsions may be lyophilized.    -   An emulsion of squalene, poloxamer 105 and Abil-Care [94]. The        final concentration (weight) of these components in adjuvanted        vaccines are 5% squalene, 4% poloxamer 105 (pluronic polyol) and        2% Abil-Care 85 (Bis-PEG/PPG-16/16 PEG/PPG-16/16 dimethicone;        caprylic/capric triglyceride).    -   An emulsion having from 0.5-50% of an oil, 0.1-10% of a        phospholipid, and 0.05-5% of a non-ionic surfactant. As        described in reference 95, preferred phospholipid components are        phosphatidylcholine, phosphatidylethanolamine,        phosphatidylserine, phosphatidylinositol, phosphatidylglycerol,        phosphatidic acid, sphingomyelin and cardiolipin. Submicron        droplet sizes are advantageous.    -   A submicron oil-in-water emulsion of a non-metabolisable oil        (such as light mineral oil) and at least one surfactant (such as        lecithin, Tween 80 or Span 80). Additives may be included, such        as QuilA saponin, cholesterol, a saponin-lipophile conjugate        (such as GPI-0100, described in reference 96, produced by        addition of aliphatic amine to desacylsaponin via the carboxyl        group of glucuronic acid), dimethyldioctadecylammonium bromide        and/or N,N-dioctadecyl-N,N-bis (2-hydroxyethyl)propanediamine.    -   An emulsion in which a saponin (e.g. QuilA or QS21) and a sterol        (e.g. a cholesterol) are associated as helical micelles [97].    -   An emulsion comprising a mineral oil, a non-ionic lipophilic        ethoxylated fatty alcohol, and a non-ionic hydrophilic        surfactant (e.g. an ethoxylated fatty alcohol and/or        polyoxyethylene-polyoxypropylene block copolymer) [98].    -   An emulsion comprising a mineral oil, a non-ionic hydrophilic        ethoxylated fatty alcohol, and a non-ionic lipophilic surfactant        (e.g. an ethoxylated fatty alcohol and/or        polyoxyethylene-polyoxypropylene block copolymer) [98].

In some embodiments an emulsion may be mixed with antigenextemporaneously, at the time of delivery, and thus the adjuvant andantigen may be kept separately in a packaged or distributed vaccine,ready for final formulation at the time of use. In other embodiments anemulsion is mixed with antigen during manufacture, and thus thecomposition is packaged in a liquid adjuvanted form. The antigen willgenerally be in an aqueous form, such that the vaccine is finallyprepared by mixing two liquids. The volume ratio of the two liquids formixing can vary (e.g. between 5:1 and 1:5) but is generally about 1:1.Where concentrations of components are given in the above descriptionsof specific emulsions, these concentrations are typically for anundiluted composition, and the concentration after mixing with anantigen solution will thus decrease.

Where a composition includes a tocopherol, any of the α, β, γ, δ, ε or ξtocopherols can be used, but α-tocopherols are preferred. The tocopherolcan take several forms e.g. different salts and/or isomers. Saltsinclude organic salts, such as succinate, acetate, nicotinate, etc.D-α-tocopherol and DL-α-tocopherol can both be used. Tocopherols areadvantageously included in vaccines for use in elderly patients (e.g.aged 60 years or older) because vitamin E has been reported to have apositive effect on the immune response in this patient group [99]. Theyalso have antioxidant properties that may help to stabilize theemulsions [100]. A preferred α-tocopherol is DL-α-tocopherol, and thepreferred salt of this tocopherol is the succinate. The succinate salthas been found to cooperate with TNF-related ligands in vivo.

C. Saponin Formulations [Chapter 22 of Ref 82]

Saponin formulations may also be used as adjuvants in the invention.Saponins are a heterogeneous group of sterol glycosides and triterpenoidglycosides that are found in the bark, leaves, stems, roots and evenflowers of a wide range of plant species. Saponin from the bark of theQuillaia saponaria Molina tree have been widely studied as adjuvants.Saponin can also be commercially obtained from Smilax ornata(sarsaprilla), Gypsophilla paniculata (brides veil), and Saponariaofficianalis (soap root). Saponin adjuvant formulations include purifiedformulations, such as QS21, as well as lipid formulations, such asISCOMs. QS21 is marketed as Stimulon™.

Saponin compositions have been purified using HPLC and RP-HPLC. Specificpurified fractions using these techniques have been identified,including QS7, QS17, QS18, QS21, QH-A, QH-B and QH-C. Preferably, thesaponin is QS21. A method of production of QS21 is disclosed in ref.101. Saponin formulations may also comprise a sterol, such ascholesterol [102].

Combinations of saponins and cholesterols can be used to form uniqueparticles called immunostimulating complexes (ISCOMs) [chapter 23 ofref. 82]. ISCOMs typically also include a phospholipid such asphosphatidylethanolamine or phosphatidylcholine. Any known saponin canbe used in ISCOMs. Preferably, the ISCOM includes one or more of QuilA,QHA & QHC. ISCOMs are further described in refs. 102-104. Optionally,the ISCOMS may be devoid of additional detergent [105].

A review of the development of saponin based adjuvants can be found inrefs. 106 & 107.

D. Virosomes and Virus-Like Particles

Virosomes and virus-like particles (VLPs) can also be used as adjuvantsin the invention. These structures generally contain one or moreproteins from a virus optionally combined or formulated with aphospholipid. They are generally non-pathogenic, non-replicating andgenerally do not contain any of the native viral genome. The viralproteins may be recombinantly produced or isolated from whole viruses.These viral proteins suitable for use in virosomes or VLPs includeproteins derived from influenza virus (such as HA or NA), Hepatitis Bvirus (such as core or capsid proteins), Hepatitis E virus, measlesvirus, Sindbis virus, Rotavirus, Foot-and-Mouth Disease virus,Retrovirus, Norwalk virus, human Papilloma virus, HIV, RNA-phages,Qβ-phage (such as coat proteins), GA-phage, fr-phage, AP205 phage, andTy (such as retrotransposon Ty protein p1). VLPs are discussed furtherin refs. 108-113. Virosomes are discussed further in, for example, ref.114

E. Bacterial or Microbial Derivatives Adjuvants suitable for use in theinvention include bacterial or microbial derivatives such as non-toxicderivatives of enterobacterial lipopolysaccharide (LPS), Lipid Aderivatives, immunostimulatory oligonucleotides and ADP-ribosylatingtoxins and detoxified derivatives thereof.

Non-toxic derivatives of LPS include monophosphoryl lipid A (MPL) and3-O-deacylated MPL (3dMPL). 3dMPL is a mixture of 3 de-O-acylatedmonophosphoryl lipid A with 4, 5 or 6 acylated chains. A preferred“small particle” form of 3 De-O-acylated monophosphoryl lipid A isdisclosed in ref. 115. Such “small particles” of 3dMPL are small enoughto be sterile filtered through a 0.22 μm membrane [115]. Other non-toxicLPS derivatives include monophosphoryl lipid A mimics, such asaminoalkyl glucosaminide phosphate derivatives e.g. RC-529 [116,117].

Lipid A derivatives include derivatives of lipid A from Escherichia colisuch as OM-174. OM-174 is described for example in refs. 118 & 119.

Immunostimulatory oligonucleotides suitable for use as adjuvants in theinvention include nucleotide sequences containing a CpG motif (adinucleotide sequence containing an unmethylated cytosine linked by aphosphate bond to a guanosine). Double-stranded RNAs andoligonucleotides containing palindromic or poly(dG) sequences have alsobeen shown to be immunostimulatory.

The CpG's can include nucleotide modifications/analogs such asphosphorothioate modifications and can be double-stranded orsingle-stranded. References 120, 121 and 122 disclose possible analogsubstitutions e.g. replacement of guanosine with2′-deoxy-7-deazaguanosine. The adjuvant effect of CpG oligonucleotidesis further discussed in refs. 123-128.

The CpG sequence may be directed to TLR9, such as the motif GTCGTT orTTCGTT [129]. The CpG sequence may be specific for inducing a Th1 immuneresponse, such as a CpG-A ODN, or it may be more specific for inducing aB cell response, such a CpG-B ODN. CpG-A and CpG-B ODNs are discussed inrefs. 130-132. Preferably, the CpG is a CpG-A ODN.

Preferably, the CpG oligonucleotide is constructed so that the 5′ end isaccessible for receptor recognition. Optionally, two CpG oligonucleotidesequences may be attached at their 3′ ends to form “immunomers”. See,for example, refs. 129 & 133-135.

A useful CpG adjuvant is CpG7909, also known as ProMune™ (ColeyPharmaceutical Group, Inc.). Another is CpG1826. As an alternative, orin addition, to using CpG sequences, TpG sequences can be used [136],and these oligonucleotides may be free from unmethylated CpG motifs. Theimmunostimulatory oligonucleotide may be pyrimidine-rich. For example,it may comprise more than one consecutive thymidine nucleotide (e.g.TTTT, as disclosed in ref. 136), and/or it may have a nucleotidecomposition with >25% thymidine (e.g. >35%, >40%, >50%, >60%, >80%,etc.). For example, it may comprise more than one consecutive cytosinenucleotide (e.g. CCCC, as disclosed in ref. 136), and/or it may have anucleotide composition with >25% cytosine(e.g. >35%, >40%, >50%, >60%, >80%, etc.). These oligonucleotides may befree from unmethylated CpG motifs. Immunostimulatory oligonucleotideswill typically comprise at least 20 nucleotides. They may comprise fewerthan 100 nucleotides.

A particularly useful adjuvant based around immunostimulatoryoligonucleotides is known as IC-31™ [137]. Thus an adjuvant used withthe invention may comprise a mixture of (i) an oligonucleotide (e.g.between 15-40 nucleotides) including at least one (and preferablymultiple) CpI motifs (i.e. a cytosine linked to an inosine to form adinucleotide), and (ii) a polycationic polymer, such as an oligopeptide(e.g. between 5-20 amino acids) including at least one (and preferablymultiple) Lys-Arg-Lys tripeptide sequence(s). The oligonucleotide may bea deoxynucleotide comprising 26-mer sequence 5′-(IC)₁₃-3′ (SEQ ID NO:175). The polycationic polymer may be a peptide comprising 11-mer aminoacid sequence KLKLLLLLKLK (SEQ ID NO: 176). The oligonucleotide andpolymer can form complexes e.g. as disclosed in references 138 & 139.

Bacterial ADP-ribosylating toxins and detoxified derivatives thereof maybe used as adjuvants in the invention. Preferably, the protein isderived from E. coli (E. coli heat labile enterotoxin “LT”), cholera(“CT”), or pertussis (“PT”). The use of detoxified ADP-ribosylatingtoxins as mucosal adjuvants is described in ref. 140 and as parenteraladjuvants in ref. 141. The toxin or toxoid is preferably in the form ofa holotoxin, comprising both A and B subunits. Preferably, the A subunitcontains a detoxifying mutation; preferably the B subunit is notmutated. Preferably, the adjuvant is a detoxified LT mutant such asLT-K63, LT-R72, and LT-G192. The use of ADP-ribosylating toxins anddetoxified derivatives thereof, particularly LT-K63 and LT-R72, asadjuvants can be found in refs. 142-149. A useful CT mutant is orCT-E29H [150]. Numerical reference for amino acid substitutions ispreferably based on the alignments of the A and B subunits ofADP-ribosylating toxins set forth in ref. 151, specifically incorporatedherein by reference in its entirety.

F. Human Immunomodulators

Human immunomodulators suitable for use as adjuvants in the inventioninclude cytokines, such as interleukins (e.g. IL-1, IL-2, IL-4, IL-5,IL-6, IL-7, IL-12 [152], etc.) [153], interferons (e.g. interferon-γ),macrophage colony stimulating factor, and tumor necrosis factor. Apreferred immunomodulator is IL-12.

G. Bioadhesives and Mucoadhesives

Bioadhesives and mucoadhesives may also be used as adjuvants in theinvention. Suitable bioadhesives include esterified hyaluronic acidmicrospheres [154] or mucoadhesives such as cross-linked derivatives ofpoly(acrylic acid), polyvinyl alcohol, polyvinyl pyrollidone,polysaccharides and carboxymethylcellulose. Chitosan and derivativesthereof may also be used as adjuvants in the invention [155].

H. Microparticles

Microparticles may also be used as adjuvants in the invention.Microparticles (i.e. a particle of ˜100 nm to ˜150 μm in diameter, morepreferably ˜200 nm to ˜30 μm in diameter, and most preferably ˜500 nm to˜10 μm in diameter) formed from materials that are biodegradable andnon-toxic (e.g. a poly(α-hydroxy acid), a polyhydroxybutyric acid, apolyorthoester, a polyanhydride, a polycaprolactone, etc.), withpoly(lactide-co-glycolide) are preferred, optionally treated to have anegatively-charged surface (e.g. with SDS) or a positively-chargedsurface (e.g. with a cationic detergent, such as CTAB).

I. Liposomes (Chapters 13 & 14 of Ref 82)

Examples of liposome formulations suitable for use as adjuvants aredescribed in refs. 156-158.

J. Polyoxyethylene Ether and Polyoxyethylene Ester Formulations

Adjuvants suitable for use in the invention include polyoxyethyleneethers and polyoxyethylene esters [159]. Such formulations furtherinclude polyoxyethylene sorbitan ester surfactants in combination withan octoxynol [160] as well as polyoxyethylene alkyl ethers or estersurfactants in combination with at least one additional non-ionicsurfactant such as an octoxynol [161]. Preferred polyoxyethylene ethersare selected from the following group: polyoxyethylene-9-lauryl ether(laureth 9), polyoxyethylene-9-steoryl ether, polyoxytheylene-8-steorylether, polyoxyethylene-4-lauryl ether, polyoxyethylene-35-lauryl ether,and polyoxyethylene-23-lauryl ether.

K. Phosphazenes

A phosphazene, such as poly[di(carboxylatophenoxy)phosphazene] (“PCPP”)as described, for example, in references 162 and 163, may be used.

L. Muramyl Peptides

Examples of muramyl peptides suitable for use as adjuvants in theinvention include N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP),N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP), andN-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1′-2′-dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamineMTP-PE).

M. Imidazoquinolone Compounds.

Examples of imidazoquinolone compounds suitable for use adjuvants in theinvention include Imiquimod (“R-837”) [164,165], Resiquimod (“R-848”)[166], and their analogs; and salts thereof (e.g. the hydrochloridesalts). Further details about immunostimulatory imidazoquinolines can befound in references 167 to 171.

N. Substituted Ureas

Substituted ureas useful as adjuvants include compounds of formula I, IIor III, or salts thereof:

-   -   as defined in reference 172, such as ‘ER 803058’, ‘ER 803732’,        ‘ER 804053’, ER 804058′, ‘ER 804059’, ‘ER 804442’, ‘ER 804680’,        ‘ER 804764’, ER 803022 or ‘ER 804057’ e.g.:

O. Further Adjuvants

Further adjuvants that may be used with the invention include:

-   -   An aminoalkyl glucosaminide phosphate derivative, such as RC-529        [173,174].

Cyclic diguanylate (‘c-di-GMP’), which has been reported as a usefuladjuvant for S. aureus vaccines [175].

A thiosemicarbazone compound, such as those disclosed in reference 176.Methods of formulating, manufacturing, and screening for activecompounds are also described in reference 176. The thiosemicarbazonesare particularly effective in the stimulation of human peripheral bloodmononuclear cells for the production of cytokines, such as TNF-α.

A tryptanthrin compound, such as those disclosed in reference 177.Methods of formulating, manufacturing, and screening for activecompounds are also described in reference 177. The thiosemicarbazonesare particularly effective in the stimulation of human peripheral bloodmononuclear cells for the production of cytokines, such as TNF-α.

-   -   A nucleoside analog, such as: (a) Isatorabine (ANA-245;        7-thia-8-oxoguanosine):

-   -   and prodrugs thereof; (b) ANA975; (c) ANA-025-1; (d) ANA380; (e)        the compounds disclosed in references 178 to 180 Loxoribine        (7-allyl-8-oxoguanosine) [181].    -   Compounds disclosed in reference 182, including: Acylpiperazine        compounds, Indoledione compounds, Tetrahydraisoquinoline (THIQ)        compounds, Benzocyclodione compounds, Aminoazavinyl compounds,        Aminobenzimidazole quinolinone (ABIQ) compounds [183,184],        Hydrapthalamide compounds, Benzophenone compounds, Isoxazole        compounds, Sterol compounds, Quinazilinone compounds, Pyrrole        compounds [185], Anthraquinone compounds, Quinoxaline compounds,        Triazine compounds, Pyrazalopyrimidine compounds, and Benzazole        compounds [186].    -   Compounds containing lipids linked to a phosphate-containing        acyclic backbone, such as the TLR4 antagonist E5564 [187,188]:    -   A polyoxidonium polymer [189,190] or other N-oxidized        polyethylene-piperazine derivative.    -   Methyl inosine 5′-monophosphate (“MIMP”) [191].    -   A polyhydroxlated pyrrolizidine compound [192], such as one        having formula:

-   -   where R is selected from the group comprising hydrogen, straight        or branched, unsubstituted or substituted, saturated or        unsaturated acyl, alkyl (e.g. cycloalkyl), alkenyl, alkynyl and        aryl groups, or a pharmaceutically acceptable salt or derivative        thereof. Examples include, but are not limited to: casuarine,        casuarine-6-α-D-glucopyranose, 3-epi-casuarine, 7-epi-casuarine,        3,7-diepi-casuarine, etc.    -   A CD1d ligand, such as an α-glycosylceramide [193-200] (e.g.        α-galactosylceramide), phytosphingosine-containing        α-glycosylceramides, OCH, KRN7000        [(2S,3S,4R)-1-O-(α-D-galactopyranosyl)-2-(N-hexacosanoylamino)-1,3,4-octadecanetriol],        CRONY-101, 3″-O-sulfo-galactosylceramide, etc.    -   A gamma inulin [201] or derivative thereof, such as algammulin.

Adjuvant Combinations

The invention may also comprise combinations of one or more of theadjuvants identified above. For example, the following adjuvantcompositions may be used in the invention: (1) a saponin and anoil-in-water emulsion [202]; (2) a saponin (e.g. QS21)+a non-toxic LPSderivative (e.g. 3dMPL) [203]; (3) a saponin (e.g. QS21)+a non-toxic LPSderivative (e.g. 3dMPL)+a cholesterol; (4) a saponin (e.g.QS21)+3dMPL+IL-12 (optionally +a sterol) [204]; (5) combinations of3dMPL with, for example, QS21 and/or oil-in-water emulsions [205]; (6)SAF, containing 10% squalane, 0.4% Tween 80™, 5% pluronic-block polymerL121, and thr-MDP, either microfluidized into a submicron emulsion orvortexed to generate a larger particle size emulsion. (7) Ribi™ adjuvantsystem (RAS), (Ribi Immunochem) containing 2% squalene, 0.2% Tween 80,and one or more bacterial cell wall components from the group consistingof monophosphorylipid A (MPL), trehalose dimycolate (TDM), and cell wallskeleton (CWS), preferably MPL+CWS (Detox™); and (8) one or more mineralsalts (such as an aluminum salt)+a non-toxic derivative of LPS (such as3dMPL).

Other substances that act as immunostimulating agents are disclosed inchapter 7 of ref. 82.

The use of an aluminium hydroxide and/or aluminium phosphate adjuvant isparticularly preferred, and antigens are generally adsorbed to thesesalts. Calcium phosphate is another preferred adjuvant. Other preferredadjuvant combinations include combinations of Th1 and Th2 adjuvants suchas CpG & alum or resiquimod & alum. A combination of aluminium phosphateand 3dMPL may be used.

The compositions of the invention may elicit both a cell mediated immuneresponse as well as a humoral immune response. This immune response willpreferably induce long lasting (e.g. neutralising) antibodies and a cellmediated immunity that can quickly respond upon exposure topnuemococcus.

Two types of T cells, CD4 and CD8 cells, are generally thought necessaryto initiate and/or enhance cell mediated immunity and humoral immunity.CD8 T cells can express a CD8 co-receptor and are commonly referred toas Cytotoxic T lymphocytes (CTLs). CD8 T cells are able to recognized orinteract with antigens displayed on MHC Class I molecules.

CD4 T cells can express a CD4 co-receptor and are commonly referred toas T helper cells. CD4 T cells are able to recognize antigenic peptidesbound to MHC class II molecules. Upon interaction with a MHC class IImolecule, the CD4 cells can secrete factors such as cytokines. Thesesecreted cytokines can activate B cells, cytotoxic T cells, macrophages,and other cells that participate in an immune response. Helper T cellsor CD4+ cells can be further divided into two functionally distinctsubsets: TH1 phenotype and TH2 phenotypes which differ in their cytokineand effector function.

Activated TH1 cells enhance cellular immunity (including an increase inantigen-specific CTL production) and are therefore of particular valuein responding to intracellular infections. Activated TH1 cells maysecrete one or more of IL-2, IFN-γ, and TNF-β. A TH1 immune response mayresult in local inflammatory reactions by activating macrophages, NK(natural killer) cells, and CD8 cytotoxic T cells (CTLs). A TH1 immuneresponse may also act to expand the immune response by stimulatinggrowth of B and T cells with IL-12. TH1 stimulated B cells may secreteIgG2a.

Activated TH2 cells enhance antibody production and are therefore ofvalue in responding to extracellular infections. Activated TH2 cells maysecrete one or more of IL-4, IL-5, IL-6, and IL-10. A TH2 immuneresponse may result in the production of IgG1, IgE, IgA and memory Bcells for future protection.

An enhanced immune response may include one or more of an enhanced TH1immune response and a TH2 immune response.

A TH1 immune response may include one or more of an increase in CTLs, anincrease in one or more of the cytokines associated with a TH1 immuneresponse (such as IL-2, IFN-γ, and TNF-(3), an increase in activatedmacrophages, an increase in NK activity, or an increase in theproduction of IgG2a. Preferably, the enhanced TH1 immune response willinclude an increase in IgG2a production.

A TH1 immune response may be elicited using a TH1 adjuvant. A TH1adjuvant will generally elicit increased levels of IgG2a productionrelative to immunization of the antigen without adjuvant. TH1 adjuvantssuitable for use in the invention may include for example saponinformulations, virosomes and virus like particles, non-toxic derivativesof enterobacterial lipopolysaccharide (LPS), immunostimulatoryoligonucleotides. Immunostimulatory oligonucleotides, such asoligonucleotides containing a CpG motif, are preferred TH1 adjuvants foruse in the invention.

A TH2 immune response may include one or more of an increase in one ormore of the cytokines associated with a TH2 immune response (such asIL-4, IL-5, IL-6 and IL-10), or an increase in the production of IgG1,IgE, IgA and memory B cells. Preferably, the enhanced TH2 immuneresponse will include an increase in IgG1 production.

A TH2 immune response may be elicited using a TH2 adjuvant. A TH2adjuvant will generally elicit increased levels of IgG1 productionrelative to immunization of the antigen without adjuvant. TH2 adjuvantssuitable for use in the invention include, for example, mineralcontaining compositions, oil-emulsions, and ADP-ribosylating toxins anddetoxified derivatives thereof. Mineral containing compositions, such asaluminium salts are preferred TH2 adjuvants for use in the invention.

Preferably, the invention includes a composition comprising acombination of a TH1 adjuvant and a TH2 adjuvant. Preferably, such acomposition elicits an enhanced TH1 and an enhanced TH2 response, i.e.,an increase in the production of both IgG1 and IgG2a production relativeto immunization without an adjuvant. Still more preferably, thecomposition comprising a combination of a TH1 and a TH2 adjuvant elicitsan increased TH1 and/or an increased TH2 immune response relative toimmunization with a single adjuvant (i.e., relative to immunization witha TH1 adjuvant alone or immunization with a TH2 adjuvant alone).

The immune response may be one or both of a TH1 immune response and aTH2 response. Preferably, immune response provides for one or both of anenhanced TH1 response and an enhanced TH2 response.

The enhanced immune response may be one or both of a systemic and amucosal immune response. Preferably, the immune response provides forone or both of an enhanced systemic and an enhanced mucosal immuneresponse. Preferably the mucosal immune response is a TH2 immuneresponse. Preferably, the mucosal immune response includes an increasein the production of IgA.

S. aureus infections can affect various areas of the body and so thecompositions of the invention may be prepared in various forms. Forexample, the compositions may be prepared as injectables, either asliquid solutions or suspensions. Solid forms suitable for solution in,or suspension in, liquid vehicles prior to injection can also beprepared (e.g. a lyophilised composition or a spray-freeze driedcomposition). The composition may be prepared for topical administratione.g. as an ointment, cream or powder. The composition may be preparedfor oral administration e.g. as a tablet or capsule, as a spray, or as asyrup (optionally flavoured). The composition may be prepared forpulmonary administration e.g. as an inhaler, using a fine powder or aspray. The composition may be prepared as a suppository or pessary. Thecomposition may be prepared for nasal, aural or ocular administratione.g. as drops. The composition may be in kit form, designed such that acombined composition is reconstituted just prior to administration to apatient. Such kits may comprise one or more antigens in liquid form andone or more lyophilised antigens.

Where a composition is to be prepared extemporaneously prior to use(e.g. where a component is presented in lyophilised form) and ispresented as a kit, the kit may comprise two vials, or it may compriseone ready-filled syringe and one vial, with the contents of the syringebeing used to reactivate the contents of the vial prior to injection.

Immunogenic compositions used as vaccines comprise an immunologicallyeffective amount of antigen(s), as well as any other components, asneeded. By ‘immunologically effective amount’, it is meant that theadministration of that amount to an individual, either in a single doseor as part of a series, is effective for treatment or prevention. Thisamount varies depending upon the health and physical condition of theindividual to be treated, age, the taxonomic group of individual to betreated (e.g. non-human primate, primate, etc.), the capacity of theindividual's immune system to synthesise antibodies, the degree ofprotection desired, the formulation of the vaccine, the treatingdoctor's assessment of the medical situation, and other relevantfactors. It is expected that the amount will fall in a relatively broadrange that can be determined through routine trials. Where more than oneantigen is included in a composition then two antigens may be present atthe same dose as each other or at different doses.

As mentioned above, a composition may include a temperature protectiveagent, and this component may be particularly useful in adjuvantedcompositions (particularly those containing a mineral adjuvant, such asan aluminium salt). As described in reference 206, a liquid temperatureprotective agent may be added to an aqueous vaccine composition to lowerits freezing point e.g. to reduce the freezing point to below 0° C. Thusthe composition can be stored below 0° C., but above its freezing point,to inhibit thermal breakdown. The temperature protective agent alsopermits freezing of the composition while protecting mineral saltadjuvants against agglomeration or sedimentation after freezing andthawing, and may also protect the composition at elevated temperaturese.g. above 40° C. A starting aqueous vaccine and the liquid temperatureprotective agent may be mixed such that the liquid temperatureprotective agent forms from 1-80% by volume of the final mixture.Suitable temperature protective agents should be safe for humanadministration, readily miscible/soluble in water, and should not damageother components (e.g. antigen and adjuvant) in the composition.Examples include glycerin, propylene glycol, and/or polyethylene glycol(PEG). Suitable PEGs may have an average molecular weight ranging from200-20,000 Da. In a preferred embodiment, the polyethylene glycol canhave an average molecular weight of about 300 Da (‘PEG-300’).

The invention provides an immunogenic composition comprising: (i) one ormore antigen(s) selected from the first, second, third or fourth antigengroups; and (ii) a temperature protective agent. This composition may beformed by mixing (i) an aqueous composition comprising one or moreantigen(s) selected from the first, second, third or fourth antigengroups, with (ii) a temperature protective agent. The mixture may thenbe stored e.g. below 0° C., from 0-20° C., from 20-35° C., from 35-55°C., or higher. It may be stored in liquid or frozen form. The mixturemay be lyophilised. The composition may alternatively be formed bymixing (i) a dried composition comprising one or more antigen(s)selected from the first, second, third or fourth antigen groups, with(ii) a liquid composition comprising the temperature protective agent.Thus component (ii) can be used to reconstitute component (i).

Methods of Treatment, and Administration of the Vaccine

The invention also provides a method for raising an immune response in amammal comprising the step of administering an effective amount of acomposition of the invention. The immune response is preferablyprotective and preferably involves antibodies and/or cell-mediatedimmunity. The method may raise a booster response.

The invention also provides at least two antigens of the invention forcombined use as a medicament e.g. for use in raising an immune responsein a mammal.

The invention also provides the use of at least two antigens of theinvention in the manufacture of a medicament for raising an immuneresponse in a mammal.

By raising an immune response in the mammal by these uses and methods,the mammal can be protected against S. aureus infection, including anosocomial infection. More particularly, the mammal may be protectedagainst a skin infection, pneumonia, meningitis, osteomyelitisendocarditis, toxic shock syndrome, and/or septicaemia.

The invention also provides a kit comprising a first component and asecond component wherein neither the first component nor the secondcomponent is a composition of the invention as described above, butwherein the first component and the second component can be combined toprovide a composition of the invention as described above. The kit mayfurther include a third component comprising one or more of thefollowing: instructions, syringe or other delivery device, adjuvant, orpharmaceutically acceptable formulating solution.

The invention also provides a delivery device pre-filled with animmunogenic composition of the invention.

The mammal is preferably a human. Where the vaccine is for prophylacticuse, the human is preferably a child (e.g. a toddler or infant) or ateenager; where the vaccine is for therapeutic use, the human ispreferably a teenager or an adult. A vaccine intended for children mayalso be administered to adults e.g. to assess safety, dosage,immunogenicity, etc. Other mammals which can usefully be immunisedaccording to the invention are cows, dogs, horses, and pigs.

One way of checking efficacy of therapeutic treatment involvesmonitoring S. aureus infection after administration of the compositionsof the invention. One way of checking efficacy of prophylactic treatmentinvolves monitoring immune responses, systemically (such as monitoringthe level of IgG1 and IgG2a production) and/or mucosally (such asmonitoring the level of IgA production), against the antigens in thecompositions of the invention after administration of the composition.Typically, antigen-specific serum antibody responses are determinedpost-immunisation but pre-challenge whereas antigen-specific mucosalantibody responses are determined post-immunisation and post-challenge.

Another way of assessing the immunogenicity of the compositions of thepresent invention is to express the proteins recombinantly for screeningpatient sera or mucosal secretions by immunoblot and/or microarrays. Apositive reaction between the protein and the patient sample indicatesthat the patient has mounted an immune response to the protein inquestion. This method may also be used to identify immunodominantantigens and/or epitopes within antigens.

The efficacy of vaccine compositions can also be determined in vivo bychallenging animal models of S. aureus infection, e.g., guinea pigs ormice, with the vaccine compositions. In particular, there are threeuseful animal models for the study of S. aureus infectious disease,namely: (i) the murine abscess model [207], (ii) the murine lethalinfection model [207] and (iii) the murine pneumonia model [208]. Theabscess model looks at abscesses in mouse kidneys after intravenouschallenge. The lethal infection model looks at the number of mice whichsurvive after being infected by a normally-lethal dose of S. aureus bythe intravenous or intraperitoneal route. The pneumonia model also looksat the survival rate, but uses intranasal infection. A useful vaccinemay be effective in one or more of these models. For instance, for someclinical situations it may be desirable to protect against pneumonia,without needing to prevent hematic spread or to promote opsonisation; inother situations the main desire may be to prevent hematic spread.Different antigens, and different antigen combinations, may contributeto different aspects of an effective vaccine.

Compositions of the invention will generally be administered directly toa patient. Direct delivery may be accomplished by parenteral injection(e.g. subcutaneously, intraperitoneally, intravenously, intramuscularly,or to the interstitial space of a tissue), or mucosally, such as byrectal, oral (e.g. tablet, spray), vaginal, topical, transdermal ortranscutaneous, intranasal, ocular, aural, pulmonary or other mucosaladministration.

The invention may be used to elicit systemic and/or mucosal immunity,preferably to elicit an enhanced systemic and/or mucosal immunity.

Preferably the enhanced systemic and/or mucosal immunity is reflected inan enhanced TH1 and/or TH2 immune response. Preferably, the enhancedimmune response includes an increase in the production of IgG1 and/orIgG2a and/or IgA.

Dosage can be by a single dose schedule or a multiple dose schedule.Multiple doses may be used in a primary immunisation schedule and/or ina booster immunisation schedule. In a multiple dose schedule the variousdoses may be given by the same or different routes e.g. a parenteralprime and mucosal boost, a mucosal prime and parenteral boost, etc.Multiple doses will typically be administered at least 1 week apart(e.g. about 2 weeks, about 3 weeks, about 4 weeks, about 6 weeks, about8 weeks, about 10 weeks, about 12 weeks, about 16 weeks, etc.).

Vaccines prepared according to the invention may be used to treat bothchildren and adults. Thus a human patient may be less than 1 year old,1-5 years old, 5-15 years old, 15-55 years old, or at least 55 yearsold. Preferred patients for receiving the vaccines are the elderly (e.g.≧50 years old, ≧60 years old, and preferably ≧65 years), the young (e.g.≦5 years old), hospitalised patients, healthcare workers, armed serviceand military personnel, pregnant women, the chronically ill, orimmunodeficient patients. The vaccines are not suitable solely for thesegroups, however, and may be used more generally in a population.

Vaccines produced by the invention may be administered to patients atsubstantially the same time as (e.g. during the same medicalconsultation or visit to a healthcare professional or vaccinationcentre) other vaccines e.g. at substantially the same time as aninfluenza vaccine, a measles vaccine, a mumps vaccine, a rubellavaccine, a MMR vaccine, a varicella vaccine, a MMRV vaccine, adiphtheria vaccine, a tetanus vaccine, a pertussis vaccine, a DTPvaccine, a conjugated H. influenzae type b vaccine, an inactivatedpoliovirus vaccine, a hepatitis B virus vaccine, a meningococcalconjugate vaccine (such as a tetravalent A-C-W135-Y vaccine), arespiratory syncytial virus vaccine, etc. Further non-staphylococcalvaccines suitable for co-administration may include one or more antigenslisted on pages 33-46 of reference 51.

Nucleic Acid Immunisation

The immunogenic compositions described above include polypeptideantigens from S. aureus. In all cases, however, the polypeptide antigenscan be replaced by nucleic acids (typically DNA) encoding thosepolypeptides, to give compositions, methods and uses based on nucleicacid immunisation. Nucleic acid immunisation is now a developed field(e.g. see references 209 to 216 etc.).

The nucleic acid encoding the immunogen is expressed in vivo afterdelivery to a patient and the expressed immunogen then stimulates theimmune system. The active ingredient will typically take the form of anucleic acid vector comprising: (i) a promoter; (ii) a sequence encodingthe immunogen, operably linked to the promoter; and optionally (iii) aselectable marker. Preferred vectors may further comprise (iv) an originof replication; and (v) a transcription terminator downstream of andoperably linked to (ii). In general, (i) & (v) will be eukaryotic and(iii) & (iv) will be prokaryotic.

Preferred promoters are viral promoters e.g. from cytomegalovirus (CMV).The vector may also include transcriptional regulatory sequences (e.g.enhancers) in addition to the promoter and which interact functionallywith the promoter. Preferred vectors include the immediate-early CMVenhancer/promoter, and more preferred vectors also include CMV intron A.The promoter is operably linked to a downstream sequence encoding animmunogen, such that expression of the immunogen-encoding sequence isunder the promoter's control.

Where a marker is used, it preferably functions in a microbial host(e.g. in a prokaryote, in a bacteria, in a yeast). The marker ispreferably a prokaryotic selectable marker (e.g. transcribed under thecontrol of a prokaryotic promoter). For convenience, typical markers areantibiotic resistance genes.

The vector of the invention is preferably an autonomously replicatingepisomal or extrachromosomal vector, such as a plasmid.

The vector of the invention preferably comprises an origin ofreplication. It is preferred that the origin of replication is active inprokaryotes but not in eukaryotes.

Preferred vectors thus include a prokaryotic marker for selection of thevector, a prokaryotic origin of replication, but a eukaryotic promoterfor driving transcription of the immunogen-encoding sequence. Thevectors will therefore (a) be amplified and selected in prokaryotichosts without polypeptide expression, but (b) be expressed in eukaryotichosts without being amplified. This arrangement is ideal for nucleicacid immunization vectors.

The vector of the invention may comprise a eukaryotic transcriptionalterminator sequence downstream of the coding sequence. This can enhancetranscription levels. Where the coding sequence does not have its own,the vector of the invention preferably comprises a polyadenylationsequence. A preferred polyadenylation sequence is from bovine growthhormone.

The vector of the invention may comprise a multiple cloning site

In addition to sequences encoding the immunogen and a marker, the vectormay comprise a second eukaryotic coding sequence. The vector may alsocomprise an IRES upstream of said second sequence in order to permittranslation of a second eukaryotic polypeptide from the same transcriptas the immunogen. Alternatively, the immunogen-coding sequence may bedownstream of an IRES.

The vector of the invention may comprise unmethylated CpG motifs e.g.unmethylated DNA sequences which have in common a cytosine preceding aguanosine, flanked by two 5′ purines and two 3′ pyrimidines. In theirunmethylated form these DNA motifs have been demonstrated to be potentstimulators of several types of immune cell.

Vectors may be delivered in a targeted way. Receptor-mediated DNAdelivery techniques are described in, for example, references 217 to222. Therapeutic compositions containing a nucleic acid are administeredin a range of about 100 ng to about 200 mg of DNA for localadministration in a gene therapy protocol. Concentration ranges of about500 ng to about 50 mg, about 1 μg to about 2 mg, about 5 μg to about 500μg, and about 20 μg to about 100 μg of DNA can also be used during agene therapy protocol. Factors such as method of action (e.g. forenhancing or inhibiting levels of the encoded gene product) and efficacyof transformation and expression are considerations which will affectthe dosage required for ultimate efficacy. Where greater expression isdesired over a larger area of tissue, larger amounts of vector or thesame amounts re-administered in a successive protocol ofadministrations, or several administrations to different adjacent orclose tissue portions may be required to effect a positive therapeuticoutcome. In all cases, routine experimentation in clinical trials willdetermine specific ranges for optimal therapeutic effect.

Vectors can be delivered using gene delivery vehicles. The gene deliveryvehicle can be of viral or non-viral origin (see generally references223 to 226).

Viral-based vectors for delivery of a desired nucleic acid andexpression in a desired cell are well known in the art. Exemplaryviral-based vehicles include, but are not limited to, recombinantretroviruses (e.g. references 227 to 237), alphavirus-based vectors(e.g. Sindbis virus vectors, Semliki forest virus (ATCC VR-67; ATCCVR-1247), Ross River virus (ATCC VR-373; ATCC VR-1246) and Venezuelanequine encephalitis virus (ATCC VR-923; ATCC VR-1250; ATCC VR 1249; ATCCVR-532); hybrids or chimeras of these viruses may also be used),poxvirus vectors (e.g. vaccinia, fowlpox, canarypox, modified vacciniaAnkara, etc.), adenovirus vectors, and adeno-associated virus (AAV)vectors (e.g. see refs. 238 to 243). Administration of DNA linked tokilled adenovirus [244] can also be employed.

Non-viral delivery vehicles and methods can also be employed, including,but not limited to, polycationic condensed DNA linked or unlinked tokilled adenovirus alone [e.g. 244], ligand-linked DNA [245], eukaryoticcell delivery vehicles cells [e.g. refs. 246 to 250] and nucleic chargeneutralization or fusion with cell membranes. Naked DNA can also beemployed. Exemplary naked DNA introduction methods are described inrefs. 251 and 252. Liposomes (e.g. immunoliposomes) that can act as genedelivery vehicles are described in refs. 253 to 257. Additionalapproaches are described in references 258 & 259.

Further non-viral delivery suitable for use includes mechanical deliverysystems such as the approach described in ref. 259. Moreover, the codingsequence and the product of expression of such can be delivered throughdeposition of photopolymerized hydrogel materials or use of ionizingradiation [e.g. refs. 260 & 261]. Other conventional methods for genedelivery that can be used for delivery of the coding sequence include,for example, use of hand-held gene transfer particle gun [262] or use ofionizing radiation for activating transferred genes [260 & 261].

Delivery DNA using PLG {poly(lactide-co-glycolide)} microparticles is aparticularly preferred method e.g. by adsorption to the microparticles,which are optionally treated to have a negatively-charged surface (e.g.treated with SDS) or a positively-charged surface (e.g. treated with acationic detergent, such as CTAB).

S. epidermidis

Although the invention focuses on S. aureus, the inventors also realisethat the sta006 and sta011 antigens have homologs in S. epidermidis. Forexample, SEQ ID NO: 234 is the ‘iron (Fe+3) ABC superfamily ATP bindingcassette transporter, binding protein’ from S. epidermidis strainM23864:W1, with 73% identity to SEQ ID NO: 42 (sta006), and SEQ ID NO:235 is the ‘putative lipoprotein’ from S. epidermidis strain RP62A, with67% identity to SEQ ID NO: 47 (sta011).

S. epidermidis is commonly present on human skin and can sometimes causeillness. Infection is usually associated with medical devices, such ascatheters, and is a cause of nosocomial infections. The resultsdisclosed herein for sta006 and sta011 against S. aureus suggest thatthe homologous proteins in S. epidermidis could be useful for immunisingagainst this pathogen.

The invention provides an immunogenic composition comprising:

-   -   (i) a polypeptide comprising an amino acid sequence: (a) having        50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%,        91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to        SEQ ID NO: 234; and/or (b) comprising a fragment of at least ‘n’        consecutive amino acids of SEQ ID NO: 234, wherein ‘n’ is 7 or        more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60,        70, 80, 90, 100, 150, 200, 250 or more);        and/or    -   (ii) a polypeptide comprising an amino acid sequence: (a) having        50% or more identity (e.g. 60%, 65%, 70%, 75%, 80%, 85%, 90%,        91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or more) to        SEQ ID NO: 235; and/or (b) comprising a fragment of at least ‘n’        consecutive amino acids of SEQ ID NO: 235, wherein ‘n’ is 7 or        more (e.g. 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 50, 60,        70, 80, 90, 100, 150, 200, 250 or more).

The composition may also include an adjuvant. These compositions areparticularly useful for immunising a mammal (including a human) againstS. epidermidis infection.

Preferred fragments of (b) comprise an epitope from SEQ ID NO: 234 or235, respectively. Other preferred fragments lack one or more aminoacids (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more) from theC-terminus and/or one or more amino acids (e.g. 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 15, 20, 25 or more) from the N-terminus of SEQ ID NO: 234/235while retaining at least one epitope of SEQ ID NO: 234/235.

More generally, the invention provides the use of the sta006 and/orsta011 homolog from any Staphylococcus species for immunising a mammalagainst that species.

Antibodies

Antibodies against S. aureus antigens can be used for passiveimmunisation. Thus the invention provides an antibody which is specificfor an antigen in the first, second, third or fourth antigen groups. Theinvention also provides the use of such antibodies in therapy. Theinvention also provides the use of such antibodies in the manufacture ofa medicament. The invention also provides a method for treating a mammalcomprising the step of administering an effective amount of an antibodyof the invention. As described above for immunogenic compositions, thesemethods and uses allow a mammal to be protected against S. aureusinfection.

The term “antibody” includes intact immunoglobulin molecules, as well asfragments thereof which are capable of binding an antigen. These includehybrid (chimeric) antibody molecules [263, 264]; F(ab′)2 and F(ab)fragments and Fv molecules; non-covalent heterodimers [265, 266];single-chain Fv molecules (sFv) [267]; dimeric and trimeric antibodyfragment constructs; minibodies [268, 269]; humanized antibody molecules[270-272]; and any functional fragments obtained from such molecules, aswell as antibodies obtained through non-conventional processes such asphage display. Preferably, the antibodies are monoclonal antibodies.Methods of obtaining monoclonal antibodies are well known in the art.Humanised or fully-human antibodies are preferred.

General

The practice of the present invention will employ, unless otherwiseindicated, conventional methods of chemistry, biochemistry, molecularbiology, immunology and pharmacology, within the skill of the art. Suchtechniques are explained fully in the literature. See, e.g., references273-280, etc.

“GI” numbering is used above. A GI number, or “GenInfo Identifier”, is aseries of digits assigned consecutively to each sequence recordprocessed by NCBI when sequences are added to its databases. The GInumber bears no resemblance to the accession number of the sequencerecord. When a sequence is updated (e.g. for correction, or to add moreannotation or information) then it receives a new GI number. Thus thesequence associated with a given GI number is never changed.

Where the invention concerns an “epitope”, this epitope may be a B-cellepitope and/or a T-cell epitope. Such epitopes can be identifiedempirically (e.g. using PEPSCAN [281,282] or similar methods), or theycan be predicted (e.g. using the Jameson-Wolf antigenic index [283],matrix-based approaches [284], MAPITOPE [285], TEPITOPE [286,287],neural networks [288], OptiMer & EpiMer [289, 290], ADEPT [291], Tsites[292], hydrophilicity [293], antigenic index [294] or the methodsdisclosed in references 295-299, etc.). Epitopes are the parts of anantigen that are recognised by and bind to the antigen binding sites ofantibodies or T-cell receptors, and they may also be referred to as“antigenic determinants”.

Where an antigen “domain” is omitted, this may involve omission of asignal peptide, of a cytoplasmic domain, of a transmembrane domain, ofan extracellular domain, etc.

The term “comprising” encompasses “including” as well as “consisting”e.g. a composition “comprising” X may consist exclusively of X or mayinclude something additional e.g. X+Y.

The term “about” in relation to a numerical value x is optional andmeans, for example, x+10%.

References to a percentage sequence identity between two amino acidsequences means that, when aligned, that percentage of amino acids arethe same in comparing the two sequences. This alignment and the percenthomology or sequence identity can be determined using software programsknown in the art, for example those described in section 7.7.18 of ref.300. A preferred alignment is determined by the Smith-Waterman homologysearch algorithm using an affine gap search with a gap open penalty of12 and a gap extension penalty of 2, BLOSUM matrix of 62. TheSmith-Waterman homology search algorithm is disclosed in ref. 301.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows bacterial counts (Log cfu/ml) after challenge of micepreviously immunised with the indicated antigens.

FIGS. 2 to 4 show survival (%) after challenge of mice previouslyimmunised with various mixtures of antigens over 14 days. In FIG. 2, thesix groups from SA-10-a are, from top to bottom at day 14. groups (i),(iii) & (iv) together, (ii), IsdB, then the negative control. In FIG. 3,the six groups from SA-10-a are, from top to bottom at day 14. groups(i), (iii) & (iv) together, (ii), IsdB, then the negative control. InFIG. 3, the six groups from SA-10-b are, from top to bottom at day 14.groups (iii), (i), (iv), (ii) and IsdB together, then the negativecontrol. In FIG. 4, the six groups from SA-14 are, from top to bottom atday 14. groups (iv), (ii), (i), (iii), negative control, and IsdB.

FIG. 5 shows collected data on mouse survival from four differentexperiments after challenge of mice previously immunised with variouscompositions (PBS negative control; IsdB antigen; and “Combo-1” and“Combo-2” antigen combinations of the invention). Individual symbolsshow the survival duration of individual mice; the horizontal bar foreach group shows the median survival duration; the percentage figuresare survival 14 days after challenge; and the p values at the top aret-Test comparisons of median survival durations between groups.

FIG. 6 shows the number of colony forming units (cfu) in mouse kidneysafter infection with 9×10⁶ cfu of Newman strain in the abscess model.Horizontal bars are averages per group, and the figure beneath eachgroup is the log reduction relative to the PBS control group.

FIG. 7 shows bacterial count (log CFU/ml) in kidneys of mice in anabscess model experiment. Mice were challenged with the followingstrains: (A) MW2; (B) LAC; (C) Staph19; or (D) MU50. Each point is anindividual animal and the bar shows the median count per group. Mice hadbeen immunised as shown on the x-axis label.

FIG. 8 shows the formation of Sta011 oligomers in the presence ofincreasing concentrations of Ca⁺⁺ ions. Numbers indicate mMconcentrations, and a * indicates the presence of 50 mM EDTA.

FIG. 9 shows IgG titers against (A) EsxAB (B) Sta006 (C) Hla-H35L (D)Sta011. Each graph has three groups, with a pair of bars per group. Theright-hand bar in a pair shows pre-immune IgG and the left-hand barshows post-immune IgG. The three groups are the compositions used forimmunising and, from left to right, are: negative control of adjuvantalone; the Combo1 combination; and the relevant antigen alone.

FIG. 10 shows bacterial counts values (log CFU/ml) in mice afterchallenge with the indicated strains. Each point is an individual animaland the bar shows the median CFU. The P value beneath the IsdB and Combocolumns is a comparison against the adjuvant-only control.

FIG. 11 shows the area of abscesses (mm²) in mice after challenge withNewman strain.

FIG. 12 shows days of survival of mice after challenge with fourdifferent strains: Newman (∘), ST-80 (□), USA300-FPR3757 (Δ) orUSA300-Lac (x) strains. Each point is an individual animal, the barshows the median survival, and the heading number shows the % of animalssurviving after 15 days. Mice received aluminium hydroxide adjuvantalone, IsdB or Combo1.

FIG. 13 shows the median survival (days) of mice after challenge. Themice had been immunised with the antigens indicated on the X-axis. Eachpoint is an individual animal and the bar shows the median survival. Theheading numbers show the % of animals surviving after 15 days.

MODES FOR CARRYING OUT THE INVENTION Antigen Selection

S. aureus proteins have been selected for use as vaccine componentsbased on various criteria.

IsdA is a surface protein involved in iron uptake. It is detectable witha high molecular weight (>250 kDa) in immunoblots of whole cell lysatesand cell wall fractions of S. aureus. Furthermore, labelled anti-IsdAantibodies revealed extracellular structures. These structures were seenin a variety of growth and infection conditions, including iron positiveconditions (in which IsdA expression is reported to be suppressed). Thestructures have a tail up to 4 μm long, with a typical orientationparallel to the mammalian cell surface. Detached IsdA-positivestructures were observed to adhere on the surface of epithelial cells,but lose cell junction localization. Epithelia/bacteria interaction maystimulate expression of the structures. In addition, the inventors havefound that IsdA is well conserved between different strains (present in36/36 strains tested; see below), thus offering protection across abroad population of circulating strains. Iron uptake is important forvirulence, so the protein is likely to be available for immune attack atpathological stages of the bacterial life cycle. The inventors havefound that the protein is not cytotoxic to human cells (see below). Theprotein can also adsorb reasonably well to aluminium hydroxide (seebelow), which is useful for stable formulation for delivery to humans.It is useful for providing an immune response to prevent hematic spreadof the bacterium.

EsxA and EsxB are small acidic dimeric secreted proteins. The inventorshave found that EsxA is highly conserved between different strains(present in 36/36 strains tested; see below), while EsxB is present in25/36 strains. The proteins are involved in persisting an infection andso are likely to be available for immune attack at pathological stagesof the bacterial life cycle. The inventors have found that a fusion ofEsxA and EsxB (‘EsxAB’) is not cytotoxic to human cells (see below). Itcan also adsorb well to aluminium hydroxide (see below), which is usefulfor stable formulation for delivery to humans. Thus the antigens areuseful for providing an immune response to prevent hematic spread of thebacterium.

Hla is a pore-forming secreted toxin. This protein is well conservedbetween different strains (present in 36/36 strains tested; see below),thus offering protection across a broad population of circulatingstrains. It is an important virulence factor so is likely to beavailable for immune attack at pathological stages of the bacterial lifecycle. It is not cytotoxic to human cells (see below). The protein canadsorb reasonably well to aluminium hydroxide (see below), which isuseful for stable formulation for delivery to humans. It is useful forproviding an immune response to prevent pneumonia.

Spa is a surface protein involved in Fc binding. The inventors havefound that this protein is well conserved between different strains(present in 36/36 strains tested), thus offering protection across abroad population of circulating strains. It is important for virulenceso is likely to be available for immune attack at pathological stages ofthe bacterial life cycle. The protein can also adsorb reasonably well toaluminium hydroxide (see below), which is useful for stable formulationfor delivery to humans. It is useful for providing an immune response toprevent hematic spread of the bacterium.

Sta006 (also known as FhuD2) is a surface protein involved in ironuptake. The inventors have found that this protein is well conservedbetween different strains (present in 36/36 strains tested; see below),thus offering protection across a broad population of circulatingstrains. The inventors have found that the protein is not cytotoxic tohuman cells (see below). The protein can also adsorb well to aluminiumhydroxide (see below), which is useful for stable formulation fordelivery to humans. It is useful for providing an immune response toprevent hematic spread of the bacterium.

Sta011 is a surface lipoprotein. The inventors have found that thisprotein is well conserved between different strains (present in 36/36strains tested; see below), thus offering protection across a broadpopulation of circulating strains. The inventors have found that theprotein is not cytotoxic to human cells (see below). The protein canalso adsorb reasonably well to aluminium hydroxide (see below), which isuseful for stable formulation for delivery to humans. It is useful forproviding an immune response to prevent hematic spread of the bacterium.This protein has been shown to assemble into oligomers in the presenceof Ca⁺⁺ ions, but not Mg⁺⁺ ions (see FIG. 8). These experiments used 5μg recombinant tag-free Sta011, incubated at 37° C. for 25 minutes withincreasing CaCl₂ concentrations from 0.5-50 mM, then analysed by gelelectrophoresis on a clear native gel. A mobility shift (indicatingoligomerisation) was evident from 2 mM Ca⁺⁺, and particularly >5 mM.These levels compare to blood Ca⁺⁺ concentrations of about 1.2 mM, serumconcentrations of about 11 mM, and milk concentrations of about 32 mM.EDTA reversed the shift.

Surface digestion [302] and/or analysis of secreted proteins revealedpeptide fragments from ClfA, ClfB, coA, eap, ebhA, ebpS, efb, emp, FnBA,FnBB, hla, IsdA, IsdB, IsdH, ukD, lukS, sdrD, sdrE, sasB, sasD, sasF,spa, sta001, sta002, sta003, sta004, sta005, sta006, sta007, sta008,sta009, sta010, sta011, sta019, sta023, sta024, sta028, sta036, sta040,sta049, sta050, sta054, sta057, sta064, sta065, sta073, sta095, sta096,sta098, sta100, sta101, sta102, sta103, sta105, sta107, sta108, sta109,sta111, sta112, sta113, sta115, sta116, sta117, sta118, sta120,NW_(—)06, NW_(—)07, NW_(—)08, NW_(—)09 and NW_(—)10 e.g. SEQ ID NOs: 228and 229 were identified as fragments of sta019.

Conjugated capsular saccharides are useful for providing opsonicimmunity. Serotypes 5 and 8 cover about 85% of clinical isolates.

Strain Coverage

A panel of 36 clinical isolates was used to represent circulatingstrains, including strains belonging to the five clonal lineagesrepresenting the vast majority of worldwide circulating CA-MRSA(community-associated methicillin-resistant S. aureus). HA-MRSA(hospital-associated MRSA) and non-MRSA strains were also included.Overall the panel included 9 HA-MRSA strains, 7 CA-MRSA strains, 2 MRSAstrains, and 18 other strains.

Genes encoding IsdA, Hla, EsxA, Sta006, Sta011, Spa, and ClfB werepresent in all 36 strains. The gene for EsxB was absent from 11/36strains, and the gene for SdrD was absent from 6/36 strains.

The encoded IsdA sequences were 95-100% identical across the panel, andthe protein was expressed in iron-limited conditions in the stationarygrowth phase. The encoded SdrD sequences were 95-100% identical in the30/36 SdrD^(+ve) panel members. The encoded EsxA sequences were 100%identical across the panel; the encoded EsxB sequences were 95-100%identical in the 25 EsxB^(+ve) strains. The encoded ClfB sequences were93-100% identical across the panel, and this protein was also found tobe highly surface-exposed in the early exponential growth phase.

Conservation in the encoded amino acid sequences were as follows (%identity):

Antigen IsdA ClfB SdrD Spa Hla EsxA EsxB Sta006 % 95-100 97-100 88-10098-100 97-100 100 95-100 99.7-100

A larger panel of 61 strains was screened for the presence of genesencoding Hla and Sta006, as well as for their expression. This panelcovered both MRSA and MSSA strains, a variety of geographical origins,and a variety of ST and clonal complex types. 9/61 strains did notexpress Hla, whereas all but one strain expressed Sta006 (data for the61st strain were inconclusive). Thus a vaccine based on Hla alone isunlikely to give adequate coverage for a universal vaccine, but thisproblem could be overcome by addition of Sta006.

Cytotoxicity and Cell Binding Studies

The analysis of the potential cellular cytotoxicity by S. aureusrecombinant antigens Hla, Hla-H35L, IsdA, IsdB, sta006, sta011 and EsxABwas conducted on HBMECs and A549 cells. Annexin V and propidium iodidestaining were used to measure the percentage of early and late apoptoticcells by flow cytometry. Endothelial cells were grown in 24 well platesup to fully confluent. Cells were then incubated for 24 hours with threedifferent concentration of recombinant antigens (10 μg/ml, 1 μg/ml, 0.1μg/ml). The combination of TNF-α and cycloheximide (CHX), which has beenreported to induce apoptosis in endothelial cells, was used as apositive control. Incubation with PBS buffer alone was a negativecontrol. Analysis was then performed by FACS.

None of the antigens induced a cytotoxic effect on HBMECs or A549 cells.Indeed, the percentage of live cell population compared to control cellsremained essentially constant up to 24 hours of incubation. In contrast,the combination of TNF-α and CHX induced a 25% increase in the number ofapoptotic cells.

HBMECs were also used as an in vitro model for testing the binding of S.aureus recombinant antigens to human endothelial cells. HBMECs weregrown up to confluence at 37° C. in humidified atmosphere in RPMI 1640medium supplemented with 10% heat-inactivated fetal calf serum, 10%NuSerum, 2 mM glutamine, 1 mM pyruvate, 1% non-essential amino acids, 1%MEM vitamins, 100 units/ml penicillin, and 100 μg/ml streptomycin.Binding of recombinant antigens to the cells was tested by indirectimmunofluorescence and analyzed by FACS. The cells positive for bindingwere measured as net mean intensity of fluorescence respect to negativecontrols, identified as unspecific antibody recognition. Bindingexperiments were performed at 4° C. Mouse polyclonal antibodies specificfor each of the recombinant antigens were used as primary antibodies andbinding was detected by R-Phycoerythrin-conjugated goat anti-mouse IgGsecondary antibody. As negative control, HBMECs were incubated withprimary polyclonal antibodies detected by fluorescence-labeled secondaryantibody or fluorescence-labeled secondary antibody alone. Binding of aknown surfaced-exposed GBS antigen was used as positive control.

Hla and Hla-H35L were the only antigens able to strongly bind toendothelial cells. The haemolytic activity of these two antigens wasalso tested.

De-fibrinated sheep and rabbit blood were used to measure theirhaemolytic activity by spectrophotometric assay. The blood was incubatedat 37° C. for 30 minutes with serial dilution 1:4 of the two proteins.Incubation with water, to cause osmotic lysis, and incubation with a S.pyogenes protein, were positive controls; as negative control, the bloodwas incubated with PBS+BSA 0.5%.

Recombinant native Hla, but not its H35L mutant form, showed haemolyticactivity on rabbit erythrocytes. The mutant was at least 150-fold lesshaemolytic than wild-type. Both proteins had no haemolytic activity onsheep blood.

Thus the S. aureus recombinant vaccine candidates do not show anycytotoxicity both on A549 epithelial cell line and HBMEC endothelialcell line. Importantly, Hla, a secreted toxin known to form pores intothe plasma membrane of host cells, could bind A549 cells but did notinduce cytotoxicity on them; it was also able to induce haemolysis ofrabbit erythrocytes. In contrast, recombinant Hla-H35L, a variant toxinwith a single amino acid substitution that cannot form cytolytic pores,did not induce cellular damage in both human cell lines and rabbiterythrocytes. These findings indicate that this mutant form of Hla maybe more safely used in a vaccine composition. None of the other antigensshowed the capacity to bind to host cells.

Adjuvant Formulation

Selected S. aureus protein antigen candidates have been formulated withaluminium hydroxide, either individually or as a combination ofproteins, with or without capsular polysaccharide conjugate(s). Theformulations have been optimized for pH and osmolarity.

The antigens were EsxA-B, Sta006, Sta011, Hla-H35L, SdrD, IsdA,IsdA₄₀₋₁₈₄, Sta019, Sta021, Sta073, ClfB₄₅₋₅₅₂, SdrD₅₃₋₅₉₂, SasF, andIsdB. These are formulated as monovalent antigens at 100 μg/ml, or ascombinations at 50 μg/ml each. Capsular saccharide conjugates from type5 or type 8 strains are added at 5 μg/ml, 10 μg/ml or 25 μg/ml.Aluminium hydroxide was used at 2 mg/ml, in a 10 mM histidine buffer (pH6.5) and with 9 mg/ml NaCl.

All monovalent and combination formulations, with or without conjugates,could be adjusted with respect to a desired pH and osmolality. Theformulations had pH in the range 6.2-7.3, and osmolality in the range248-360 mOsm/kg. Glycerol was excluded from formulations as it had anegative impact on osmolality.

All proteins tested, in various monovalent and combination formulations,adsorbed well to the aluminium hydroxide adjuvant, except for IsdA,IsdA₄₀₋₁₈₄, Sta019, and Sta073.

The individual Sta006, Sta011, EsxA-B and Hla-H35L proteins werecompletely adsorbed, and could be desorbed without altering theirpre-adsorption electrophoretic profile.

Each antigen in a combination of Sta006, Sta011, EsxA-B and Hla-H35L wascompletely adsorbed, with no inter-antigen competition for the adjuvant.The antigens in a combination of Sta006, Sta011, EsxA-B and IsdA₄₀₋₁₈₄were also completely adsorbed, except for IsdA₄₀₋₁₈₄, which behaved inthe same way as the monovalent protein. For both combinations, theantigens could be desorbed without altering their pre-adsorptionelectrophoretic profile.

The additional presence of type 5 and/or type 8 conjugates also did notchange the adsorption or desorption characteristics of the antigens e.g.in combination with Sta006+Sta011+EsxA-B.

A short stability study (2 weeks at 4° C.) was performed to evaluate thestability of monovalent formulations and to evaluate antigen integrity.All tested formulations were stable for their pH and osmolality. Allantigens remained completely adsorbed to the adjuvant. All antigensmaintained their desorption characteristics. There was no evidence ofincreased degradation or aggregation of antigens after desorption.

Efficacy Testing

Individual antigens sta006, sta011, sta012, sta017, sta019, sta021 andsta028 were tested for their ability to protect against IV challenge by1.2×10⁷ cfu of Newman strain (type 5). Results are shown in FIG. 1. Allantigens reduced bacterial numbers compared with the control, and thebest results were seen with sta006, sta011 and sta019.

Further individual antigens were tested: (i) NW_(—)10; (ii) IsdA₄₀₋₁₈₄;(iii) Sta002; (iv) Sta003; (v) Sta073; (vi) Sta101; (vii) Sta014; (viii)Hla-PSGS; (ix) SdrD_(cnaB). The increase in survival, compared to thenegative control group, 15 days after challenge was: (i) 50%; (ii) 19%;(iii) 37%; (iv) 43%; (v) 25%; (vi) 12%; (vii) 25%; (viii) 56%; (ix) 39%.

Two hybrid polypeptides were also tested: (i) HlaH35L-EsxAB; (ii)Sta006-EsxAB. The increase in survival after challenge, compared to thenegative control group, was: (i) 25%; (ii) 25%.

Table 2 gives a summary of results obtained with various antigens in theabscess model.

Experiment SA-10-a tested the efficacy of antigen combinations. Sixgroups of twelve CD-1 mice received a negative control (PBS), IsdB, orone of the following combinations, adjuvanted with aluminium hydroxide:(i) EsxAB+Hla-H35L; (ii) Sta006+Sta011+EsxAB; (iii)Sta006+Sta011+EsxAB+Hla-H35L; or (iv) Sta006+Sta011+IsdA₄₀₋₁₈₄+EsxAB.Two administrations were given, at days 0 and 14. At day 24 micereceived 3×10⁸ cfu of Newman strain staphylococcus and survival in eachgroup was assessed every 24 hours for two weeks. Results are shown inFIG. 2. After 14 days, 25% of animals in the positive control group hadsurvived, but 50% of animals in group (ii) had survived, as had 58% ofanimals in groups (iii) & (iv), and 75% in group (i).

Experiment SA-10-b used the same methods to test: (i)ClfB₄₅₋₅₅₂+Hla-H35L+Sta006+EsxAB; (ii) ClfB₄₅₋₅₅₂+Sta011+Sta006+EsxAB;(iii) ClfB₄₅₋₅₅₂+IsdA₄₀₋₁₈₄+Sta006+EsxAB; or (iv)SdrD₅₃₋₅₉₂+IsdA₄₀₋₁₈₄+Sta006+EsxAB. Results are shown in FIG. 3. After14 days, 25% of animals in the positive control group and in group (ii)had survived, but 33% of animals in group (iv) had survived, 75% ofanimals in group (i), and 83% of animals in group (iii).

Further combinations were also used to immunise mice. The combinationswere typically adjuvanted with aluminium hydroxide (see above) and wereadministered on days 0 and 14. The immunisations were in CD1 mice, 12per group. On day 24 the mice were challenged with a lethal dose of livebacteria and survival was then followed for 14 further days. Forcomparison, PBS was used as a negative control and IsdB as a positivecontrol [2].

Experiment SA-11 tested: (i) a type 5 conjugate combined withEsxAB+Sta006+Sta011; (ii) EsxAB+Sta019+Sta006+Sta011; (iii) a type 5conjugate+Hla-H35L+Sta006+Sta011; (iv) EsxAB+Hla-H35L+Sta006+Sta011; or(v) EsxAB+IsdA₄₀₋₁₈₄+Sta006+Sta011. 14 days after challenge all of thenegative control animals had died, but 42% of positive control animalshad survived. Survival results in the test groups were as follows: (i)67%; (ii) 42%; (iii) 75%; (iv) 33%; and (v) 25%.

Experiment SA-12 tested: (i) Hla-H35L+IsdA₄₀₋₁₈₄+Sta006+Sta011; (ii)Hla-H35L+EsxAB+Sta006+Sta011; (iii) EsxAB+IsdA₄₀₋₁₈₄₊Sta006+Sta011; (iv)EsxAB+IsdA+Sta006+Sta011. 14 days after challenge 8% of the negativecontrol animals and 17% of positive control animals had survived.Survival results in the test groups were as follows: (i) 50%; (ii) 50%;(iii) 25%; (iv) 33%.

Experiment SA-14 tested: (i) EsxAB+Hla-H35L+Sta006+Sta011; (ii)EsxAB+IsdA₄₀₋₁₈₄ Sta006+Sta011; (iii) Sta006+Sta011+Sta019+EsxAB; (iv)Sta006+Sta011+Sta019+Hla-H35L. 14 days after challenge with 5×10⁸ CFU ofNewman strain, 18% of the negative control animals and 9% of positivecontrol animals had survived; survival results in the test groups wereas follows: (i) 58%; (ii) 67%; (iii) 42%; (iv) 83%. Survival numbersover 14 days are shown in FIG. 4, showing that all combinationsperformed better than the two controls on every post-challenge day.

Experiment SA-17a tested: (i) EsxAB+Sta006+Sta011+serotype 5conjugate+serotype 8 conjugate; (ii) EsxAB+Sta073+Sta011+serotype 5conjugate+serotype 8 conjugate; (iii) EsxAB+Hla-H35L+Sta011+Sta073.Compared to the negative control, the increase in survival 15 days afterchallenge with Newman strain was: (i) 17%; (ii) 42%; (iii) 34%. Themedian survival in groups (ii) and (iii) was the full 15 days, and was12 days in group (i).

Further antigen combination experiments tested: (a) serotype 5conjugate+serotype 8 conjugate+EsxAB+Sta006+Sta011; (b)Sta002+Sta003+Sta021+NW-10; (c) EsxAB+HlaH35L+Sta006+Sta019; and (d)EsxAB+Sta006+Sta019. Compared to the negative control, the increase insurvival after challenge with Newman strain was: (a) 37%; (b) 36%; (c)13%; and (d) 0%.

Survival data from studies SA-10, SA-11, SA-12 and SA-14 were combinedto assess the efficacy of two combinations when compared to PBS or IsdB.“Combo-1” was EsxAB+Hla-H35L+Sta006+Sta011 (with polypeptides comprisingSEQ ID NOs: 241, 150, 246 & 247). “Combo-2” wasEsxAB+IsdA₄₀₋₁₈₄+Sta006+Sta011. The median survival times for each groupof 48 mice after 14 days were compared. Whereas the PBS and IsdB groupshad a median survival time of 1 day, mice in the “Combo-1” and “Combo-2”groups had a median survival time of 14 days. The differences in mediansurvival duration were compared by a t-test: survival in the “Combo-1”group was statistically superior to both the PBS group (p<0.0001) andthe IsdB group (p<0.0001); survival in the “Combo-2” group wasstatistically superior to both the PBS group (p<0.0001) and the IsdBgroup (p=0.0049). These data are shown in FIG. 5.

FIG. 6 shows data with Combo-1 and Combo-2 in the abscess model. Kidneysof mice are isolated after challenge and are then homogenised andplated. The cfu count indicates the level of abscess formation. FIG. 6shows data from a single experiment. The numbers beneath the data showthe log reduction relative to the PBS group. The reduction is bigger inthe two combination groups than with IsdB alone, with U-test (one tail)values of 0.0001 for Combo-1 and 0.0005 for Combo-2. The same effect wasseen in the two combination groups in a second experiment in which anIsdB group was not included.

Further experiments compared protection achieved with Combo-1, IsdB orPBS against challenge with three different strains: Staph-19,FPR3757(USA300) and Lac(USA300). There were 44 mice per group andresults were as follows (see also FIG. 12), including one-tailedp-values for the survival proportion, where: P1 compares Combo-1 withPBS; P2 compares Combo-1 with IsdB; and P3 compared PBS with IsdB:

Staph-19 FPR3757 Lac Survival % Days % Days % Days PBS 20 1 45 8 47 7IsdB 32 1 52 15 61 15 Combo-1 80 15 91 15 89 15 P1 <0.0001 — <0.0001 —0.0001 — P2 <0.0001 — <0.0004 — 0.0052 — P3 0.1715 — 0.2137 — 0.1789 —

Further experiments showed that immunisation with adjuvanted Combo1reduced CFU counts after challenge with Newman, USA100, CC30 and USA300strains, when compared to immunisation with adjuvant alone (aluminiumhydroxide) or IsdB. FIG. 10 shows CFU values (log/ml) for the fourchallenge strains. The lowest count, with p<0.015 in each case, wasachieved with Combo1. The area of abscess was also assessed and was alsolower in the Combo1-immunised mice (e.g. FIG. 11).

Further experiments showed that Combo1 is highly protective againstclinically relevant strains in the sepsis model, and always achieved ahigher survival % than IsdB. FIG. 12 shows that the median survival inCombo1-immunised mice (40 per group, 3 experiments) was the full 15 dayswhen challenged with Newman, ST-80, FPR3757 or Lac strains, and that theproportion of mice surviving was ≧75%. In contrast, the median survivalin IsdB-immunised mice was only 1 day with Newman and ST-80 challenge,with <65% survival for all four challenge strains.

Comparison of Combo1 to its Individual Polypeptides

Various tests were performed to compare Combo1 to its four individualpolypeptides (i.e. EsxAB, Hla-H35L, Sta006, Sta011), as well as to IsdBor to an antigen-free negative control.

The opsonophagocytic activity of sera from immunised animals was tested.Sera were obtained using (i) the four individual polypeptides, (ii) allpairs of the polypeptides, (iii) all triplets, or (iv) the full Combo1combination. For comparison, anti-IsdB serum was used. Pre-immune andnegative control sera showed no killing of Newman strain in this assay.In a first experiment: anti-IsdB serum showed 27% killing; sera againsteach of the four individual polypeptides showed between 26-34% killing;all multi-polypeptide combinations showed at least 34% killing; and seraraised with Combo-1 showed 39% killing. In a second experiment sera withCombo-1 showed 43% killing but anti-IsdB serum performed slightlybetter; all single or multi-polypeptide sera using the Combo-1polypeptides showed at least 26% killing.

Further experiments looked at passive protection achieved bytransferring into mice (20 per group, 8 week old CD1 mice) antiserumfrom immunised rabbits. Four groups received 200 μl of sera from rabbitsimmunised with one of EsxAB, Hla-H35L, Sta006, Sta011; a fifth groupreceived 50 μl of each serum (200 μl in total). Two other groupsreceived serum from IsdB-immunised rabbits or serum from rabbitsimmunised with saline+adjuvant. 15 minutes later the mice werechallenged intraperitoneally (10⁸ CFU of Newman strain) and thenmortality was assessed after 14 days. Results were as follows:

EsxA-B Sta006 Sta011 HlaH35L Combo1 IsdB −ve ctrl Survival 5% 26% 0% 15%25% 10% 5%

In further experiments the level of specific antibodies induced in CD1mice were examined to assess the immunogenicity of the four polypeptidesin Combo 1. Compositions included either 20 μg of each of the foursingle polypeptides, or 4×10 μg in the combination. The compositionsincluded an aluminium hydroxide adjuvant. Serum levels ofantigen-specific IgG were determined by Luminex 4Plex assay As shown inFIG. 9, all four polypeptides were highly immunogenic in CD1 mice ontheir own and in combination. In each case the titer against apolypeptide was higher when it was administered in the combination thanwhen administered alone (compare middle and right pairs).

Further experiments compared protection achieved either with Combo-1 orwith its four individual polypeptides. IsdB was also included forcomparison. The proportions of animals surviving (40 animals per group)15 days after challenge with Newman strain, and the average (median)survival in days, were as follows, including a one-tailed p-value of thesurviving proportion in comparison with a PBS+adjuvant negative control:

EsxA-B Sta006 Sta011 HlaH35L Combo1 IsdB PBS Survival 34% 28% 16% 39%59% 22% 5% p 0.0017 0.0003 0.0064 <0.0001 <0.0001 0.0006 — Days 1 2 1 1015 1 0

The murine abscess model was used to compare the four individualpolypeptides with the Combo1 combination. In some experiments mice wereimmunised with IsdB for comparison. Antigens were adjuvanted withaluminium hydroxide, and adjuvant alone was used as a negative control.FIG. 7 shows the numbers of bacteria in animals' kidneys after challengewith four different strains. The lowest average counts were seen for theCombo1 combination.

Challenge experiments were performed following immunisation with (i) thefour individual polypeptides, (ii) all pairs, (iii) all triplets, or(iv) the full Combo1 combination. IsdB or buffer alone were used forcomparison. Survival results from 24 mice per group (3 experiments)after challenge with 5×10⁸ CFU of Newman strain are shown in FIG. 13.The median survival for IsdB was only 2 days. The median survival forthe individual Combo1 polypeptides ranged from 1-6 days. Pairs of thepolypeptides gave median survival of 2-11 days. Triplets gave mediansurvival of 8-15 days. The full Combo1 combination gave a mediansurvival of the full 15 days, with 59% of mice surviving this long (cf.only 35% with IsdB).

It will be understood that the invention has been described by way ofexample only and modifications may be made whilst remaining within thescope and spirit of the invention.

TABLE 1 NOMENCLATURE CROSS-REFERENCE; Staphylococcus aureus vaccinecandidates, in silico selection. SEQ ID NCTC 8325 strain Newman strainNO Name SAOUHSC_# GI NMWN_# GI 1 clfA SAOUHSC_00812 88194572 NWMN_0756151220968 2 clfB SAOUHSC_02963 88196585 NWMN_2529 151222741 3 coASAOUHSC_00192 88194002 NWMN_0166 151220378 4 eap SAOUHSC_02161 88195840NWMN_1872 151222084 5 ebhA SAOUHSC_01447 88195168 — — 6 ebpSSAOUHSC_01501 88195217 NWMN_1389 151221601 7 efb SAOUHSC_01114 88194860NWMN_1069 151221281 8 emp SAOUHSC_00816 88194575 NWMN_0758 151220970 9esaC SAOUHSC_00264 88194069 — — 10 esxA SAOUHSC_00257 88194063 — — 11esxB SAOUHSC_00265 88194070 — — 12 FnBA SAOUHSC_02803 88196438 NWMN_2399151222611 13 FnBB SAOUHSC_02802 88196437 NWMN_2397 151222609 14 hlaSAOUHSC_01121 88194865 NWMN_1073 151221285 15 hlgB SAOUHSC_0271088196350 — — 16 hlgC SAOUHSC_02709 88196349 — — 17 isdA SAOUHSC_0108188194829 NWMN_1041 151221253 18 isdB SAOUHSC_01079 88194828 — — 19 isdCSAOUHSC_01082 88194830 — — 20 isdG SAOUHSC_01089 88194836 — — 21 isdHSAOUHSC_01843 88195542 NWMN_1624 151221836 22 isdI SAOUHSC_0013088193943 — — 23 lukD SAOUHSC_01954 88195647 NWMN_1718 151221930 24 lukESAOUHSC_01955 88195648 — — 25 lukF SAOUHSC_02241 88195914 — — 26 lukSSAOUHSC_02243 88195915 NWMN_1928 151222140 27 nuc SAOUHSC_01316 88195046— — 28 sasA SAOUHSC_02990 88196609 — — 29 sasB SAOUHSC_02404 88196065 —— 30 sasC SAOUHSC_01873 88195570 — — 31 sasD SAOUHSC_00094 88193909 — —32 sasF SAOUHSC_02982 88196601 — — 33 sdrC SAOUHSC_00544 88194324 — — 34sdrD SAOUHSC_00545 88194325 — — 35 sdrE2 — — NWMN_0525 151220737 36 spaSAOUHSC_00069 88193885 NWMN_0055 151220267 37 sta001 SAOUHSC_0002588193846 NWMN_0022 151220234 38 sta002 SAOUHSC_00356 88194155 NWMN_0364151220576 39 sta003 SAOUHSC_00400 88194195 NWMN_0401 151220613 40 sta004SAOUHSC_00749 88194514 NWMN_0705 151220917 41 sta005 SAOUHSC_0112788194870 NWMN_1077 151221289 42 sta006 SAOUHSC_02554 88196199 NWMN_2185151222397 43 sta007 SAOUHSC_02571 88196215 NWMN_2199 151222411 44 sta008SAOUHSC_02650 88196290 NWMN_2270 151222482 45 sta009 SAOUHSC_0270688196346 NWMN_2317 151222529 46 sta010 SAOUHSC_02887 88196515 NWMN_2469151222681 47 sta011 SAOUHSC_00052 88193872 — — 48 sta012 SAOUHSC_0010688193919 — — 49 sta013 SAOUHSC_00107 88193920 — — 50 sta014SAOUHSC_00137 88193950 — — 51 sta015 SAOUHSC_00170 88193980 — — 52sta016 SAOUHSC_00171 88193981 — — 53 sta017 SAOUHSC_00186 88193996 — —54 sta018 SAOUHSC_00201 88194011 — — 55 sta019 SAOUHSC_00248 88194055NWMN_0210 151220422 56 sta020 SAOUHSC_00253 88194059 — — 57 sta021SAOUHSC_00256 88194062 — — 58 sta022 SAOUHSC_00279 88194083 — — 59sta023 SAOUHSC_00284 88194087 — — 60 sta024 SAOUHSC_00300 88194101 — —61 sta025 SAOUHSC_00362 88194160 — — 62 sta026 SAOUHSC_00404 88194198 —— 63 sta027 SAOUHSC_00661 88194426 — — 64 sta028 SAOUHSC_00671 88194436NWMN_0634 151220846 65 sta029 SAOUHSC_00754 88194518 — — 66 sta030SAOUHSC_00808 88194568 — — 67 sta031 SAOUHSC_00860 88194617 — — 68sta032 SAOUHSC_00958 88194715 — — 69 sta033 SAOUHSC_00987 88194744 — —70 sta034 SAOUHSC_00988 88194745 — — 71 sta035 SAOUHSC_00998 88194754 —— 72 sta036 SAOUHSC_01084 88194831 — — 73 sta037 SAOUHSC_01085 88194832— — 74 sta038 SAOUHSC_01088 88194835 — — 75 sta039 SAOUHSC_0112488194868 — — 76 sta040 SAOUHSC_01125 88194869 NWMN_1076 151221288 77sta041 SAOUHSC_01175 88194914 — — 78 sta042 SAOUHSC_01180 88194919 — —79 sta043 SAOUHSC_01219 88194955 — — 80 sta044 SAOUHSC_01508 88195223 —— 81 sta045 SAOUHSC_01627 88195337 — — 82 sta046 SAOUHSC_01918 88195613— — 83 sta047 SAOUHSC_01920 88195615 — — 84 sta048 SAOUHSC_0194988195642 — — 85 sta049 SAOUHSC_01972 88195663 NWMN_1733 151221945 86sta050 SAOUHSC_02127 88195808 — — 87 sta051 SAOUHSC_02147 88195827 — —88 sta052 SAOUHSC_02246 88195918 — — 89 sta053 SAOUHSC_02257 88195928 —— 90 sta054 SAOUHSC_02333 88195999 — — 91 sta055 SAOUHSC_02448 88196100— — 92 sta056 SAOUHSC_02463 88196115 — — 93 sta057 SAOUHSC_0257688196220 NWMN_2203 151222415 94 sta058 SAOUHSC_02690 88196330 — — 95sta059 SAOUHSC_02708 88196348 — — 96 sta060 SAOUHSC_02767 88196403 — —97 sta061 SAOUHSC_02783 88196419 — — 98 sta062 SAOUHSC_02788 88196424 —— 99 sta063 SAOUHSC_02971 88196592 — — 100 sta064 SAOUHSC_03006 88196625NWMN_2569 151222781 101 sta065 SAOUHSC_00051 88193871 — — 102 sta066SAOUHSC_00172 88193982 — — 103 sta067 SAOUHSC_00176 88193986 — — 104sta068 SAOUHSC_00327 88194127 — — 105 sta069 SAOUHSC_00427 88194219 — —106 sta070 SAOUHSC_00773 88194535 — — 107 sta071 SAOUHSC_00854 88194612— — 108 sta072 SAOUHSC_00872 88194629 — — 109 sta073 SAOUHSC_0099488194750 NWMN_0922 151221134 110 sta074 SAOUHSC_01220 88194956 — — 111sta075 SAOUHSC_01256 88194989 — — 112 sta076 SAOUHSC_01263 88194996 — —113 sta077 SAOUHSC_01317 88195047 — — 114 sta078 SAOUHSC_01857 88195555— — 115 sta079 SAOUHSC_01935 88195630 — — 116 sta080 SAOUHSC_0193688195631 — — 170 sta081 SAOUHSC_01938 88195633 117 sta082 SAOUHSC_0193988195634 — — 118 sta083 SAOUHSC_01941 88195635 — — 119 sta084SAOUHSC_01942 88195636 — — 120 sta085 SAOUHSC_02171 88195848 — — 121sta086 SAOUHSC_02327 88195993 — — 122 sta087 SAOUHSC_02635 88196276 — —123 sta088 SAOUHSC_02844 88196477 — — 124 sta089 SAOUHSC_02855 88196486— — 125 sta090 SAOUHSC_02883 88196512 — — 126 sta091 SAOUHSC_0068588194450 — — 127 sta092 SAOUHSC_00174 88193984 — — 128 sta093SAOUHSC_01854 88195552 — — 129 sta094 SAOUHSC_01512 88195226 — — 130sta095 SAOUHSC_00383 88194180 NWMN_0388 151220600 131 sta096SAOUHSC_00384 88194181 — — 132 sta097 SAOUHSC_00386 88194182 — — 133sta098 SAOUHSC_00389 88194184 NWMN_0391 151220603 134 sta099SAOUHSC_00390 88194185 — — 135 sta100 SAOUHSC_00391 88194186 — — 136sta101 SAOUHSC_00392 88194187 NWMN_0394 151220606 137 sta102SAOUHSC_00393 88194188 — — 138 sta103 SAOUHSC_00394 88194189 — — 139sta104 SAOUHSC_00395 88194190 — — 140 sta105 SAOUHSC_00399 88194194NWMN_0400 151220612 141 sta106 SAOUHSC_01115 88194861 — — 177 sta107SAOUHSC_00354 88194153 NWMN_0362 151220574 178 sta108 SAOUHSC_0071788194482 NWMN_0677 151220889 179 sta109 SAOUHSC_02979 88196599 NWMN_2543151222755 180 sta110 SAOUHSC_01039 88194791 181 sta111 SAOUHSC_0100588194760 NWMN_0931 151221143 182 sta112 SAOUHSC_00634 88194402 NWMN_0601151220813 183 sta113 SAOUHSC_00728 88194493 NWMN_0687 151220899 184sta114 SAOUHSC_00810 88194570 185 sta115 SAOUHSC_00817 88194576NWMN_0759 151220971 186 sta116 SAOUHSC_01112 88194858 NWMN_1067151221279 187 sta117 SAOUHSC_02240 88195913 NWMN_1926 151222138 188sta118 SAOUHSC_01150 88194892 NWMN_1096 151221308 200 sta119SAOUHSC_01100 88194846 201 sta120 SAOUHSC_00365 88194163 142 NW_6 — —NWMN_0757 151220969 143 NW_9 — — NWMN_0958 151221170 144 NW_10 — —NWMN_1066 151221278 145 NW_7 — — NWMN_1876 151222088 146 NW_8 — —NWMN_1877 151222089 147 NW_2 — — NWMN_1883 151222095 148 NW_1 — —NWMN_1924 151222136 149 NW_5 — — NWMN_2392 151222604

TABLE 2 ABSCESS MODEL RESULTS SUMMARY Immunising infecting strainantigen(s) Adjuvant & dose Reduction** Fnb alum* Newman 1.4E+07 2.13Sta005 alum Newman 1.4E+07 1.26 LukE alum Newman 1.4E+07 1.68 SasD alumNewman 1.4E+07 0.10 SpA alum Newman 1.4E+07 0.41 SasFHis alum Newman1.4E+07 1.33 CoA alum Newman 1.4E+07 1.01 Sta028 alum Newman 1.2E+071.85 Sta017 alum Newman 1.2E+07 1.23 Sta006 alum Newman 1.2E+07 2.33Sta012 alum Newman 1.2E+07 1.69 Sta011 alum Newman 1.2E+07 2.66 Sta019alum Newman 1.2E+07 2.36 Sta021 alum Newman 1.2E+07 1.58 IsdA + EsxABalum Newman 1.8E+07 0.11 EsxAB alum Newman 1.8E+07 1.31 NW_1 alum Newman1.8E+07 1.00 NW_10 alum Newman 1.8E+07 −0.65 Sta073 alum Newman 1.8E+071.46 Sta002 alum Newman 1.8E+07 0.17 Sta064 alum Newman 1.8E+07 1.04Sta014 alum Newman 1.8E+07 1.74 Sta002 alum Newman 1.0E+07 0.52 Sta014alum Newman 1.0E+07 1.02 Sta064 alum Newman 1.0E+07 1.22 Sta006 alumNewman 1.0E+07 0.80 Sta073 alum Newman 1.0E+07 0.92 NW_1 alum Newman1.0E+07 0.77 NW_10 alum Newman 1.0E+07 2.25 Sta017 alum Newman 1.0E+072.13 Sta028 alum Newman 1.0E+07 0.64 Sta021 alum Newman 1.0E+07 1.03Sta019 alum Newman 1.0E+07 1.28 Sta011 alum Newman 1.0E+07 0.78 IsdBalum Newman 1.0E+07 1.22 IsdA₄₀₋₁₈₄ none Newman 1.0E+07 0.58 Sta006 noneNewman 1.0E+07 0.30 Sta011 none Newman 1.0E+07 0.62 EsxAB none Newman1.0E+07 1.09 Sasf none Newman 1.0E+07 0.11 IsdB none Newman 1.0E+07 0.93IsdA₄₀₋₁₈₄ alum Newman 1.0E+07 1.02 Sta006 alum Newman 1.0E+07 0.45Sta011 alum Newman 1.0E+07 0.80 EsxAB alum Newman 1.0E+07 0.47 Sasf alumNewman 1.0E+07 −0.78 IsdB alum Newman 1.0E+07 1.24 Type 5 conjugate +alum Newman 1.5E+07 0.34 IsdA₄₀₋₁₈₄ Type 5 conjugate alum Newman 1.5E+070.72 IsdA₄₀₋₁₈₄ alum Newman 1.5E+07 1.08 Type 5 conjugate MF59 Newman1.5E+07 0.45 IsdB alum Newman 1.5E+07 1.50 ClfB₄₅₋₅₅₂ alum Newman1.5E+07 −0.05 Sta019 alum Newman 1.5E+07 0.82 IsdA₄₀₋₁₈₄ + alum Newman1.5E+07 0.72 ClfB₄₅₋₅₅₂ Type 8 conjugate alum Becker 4.0E+07 1.51 Type 8conjugate MF59 Becker 4.0E+07 0.35 EsxAB + Sta019 + alum Newman 1.0E+071.54 Sta006 + Sta011 combo1 alum Newman 1.0E+07 2.04 EsxAB +IsdA₄₀₋₁₈₄ + alum Newman 1.0E+07 0.84 Sta006 + Sta011 SdrD₅₃₋₅₉₂ alumNewman 1.0E+07 1.15 Sta105 alum Newman 1.0E+07 0.54 Sta101 alum Newman1.0E+07 1.51 Sta116 alum Newman 1.0E+07 1.23 Sta106 alum Newman 1.0E+071.20 Sta107 alum Newman 1.0E+07 1.77 Sta004 alum Newman 1.0E+07 0.70Sta003 alum Newman 1.0E+07 1.32 EsxAB + Sta019 + alum Newman 9.0E+063.04 Sta006 + Sta011 combo1 alum Newman 9.0E+06 2.53 EsxAB +IsdA₄₀₋₁₈₄ + alum Newman 9.0E+06 1.85 Sta006 + Sta011 SdrD₅₃₋₅₉₂ alumNewman 9.0E+06 1.80 Sta105 alum Newman 9.0E+06 0.60 Sta101 alum Newman9.0E+06 0.83 Sta116 alum Newman 9.0E+06 1.96 Sta106 alum Newman 9.0E+062.56 IsdB alum Newman 9.0E+06 1.37 Sta004 alum Newman 9.0E+06 1.01Sta003 alum Newman 9.0E+06 2.20 IsdB alum Newman 1.0E+07 0.83 Sta107alum Newman 1.0E+07 0.24 SrdC₅₁₋₅₁₈ alum Newman 1.0E+07 0.84 SdrE₅₃₋₆₃₂alum Newman 1.0E+07 1.08 Hla₂₇₋₇₆ alum Newman 1.0E+07 0.18 EsxAB +HlaH35L + alum Newman 1.0E+07 0.59 Sta006 + Sta021 EsxAB + HlaH35L +alum Newman 1.0E+07 0.85 Sta006 + Sta019 EsxAB + HlaH35L + alum Newman1.0E+07 1.88 Sta006 + Sta017 EsxAB + Hla27-76 + alum Newman 1.0E+07 1.49Sta006 + Sta021 Hla₂₇₋₇₆ + Sta006 + alum Newman 1.0E+07 0.00 Sta017 +Sta019 IsdB alum Newman 1.2E+07 1.07 Sta107 alum Newman 1.2E+07 1.35SrdC₅₁₋₅₁₈ alum Newman 1.2E+07 2.17 SdrE₅₃₋₆₃₂ alum Newman 1.2E+07 2.82Hla₂₇₋₇₆ alum Newman 1.2E+07 0.17 EsxAB + HlaH35L + alum Newman 1.2E+071.70 Sta006 + Sta021 EsxAB + HlaH35L + alum Newman 1.2E+07 1.20 Sta006 +Sta019 EsxAB + HlaH35L + alum Newman 1.2E+07 1.52 Sta006 + Sta017EsxAB + Hla₂₇₋₇₆ + alum Newman 1.2E+07 1.81 Sta006 + Sta021 Hla₂₇₋₇₆ +Sta006 + alum Newman 1.2E+07 0.89 Sta017 + Sta019 IsdB alum Mu-503.8E+07 0.44 Combo1 alum Mu-50 3.8E+07 1.73 IsdB alum USA 200 2.0E+071.17 Combo1 alum USA 200 2.0E+07 1.87 IsdB alum USA 300 3.0E+07 0.09Combo1 alum USA 300 3.0E+07 2.19 IsdB alum Staph19 2.7E+07 0.66 Combo1alum Staph19 2.7E+07 0.46 IsdB alum Mu-50 4.5E+07 0.98 Combo1 alum Mu-504.5E+07 0.76 IsdB alum USA 200 1.6E+07 0.18 Combo1 alum USA 200 1.6E+070.19 IsdB alum USA 300 2.2E+07 −0.21 Combo1 alum USA 300 2.2E+07 −0.29IsdB alum Staph19 2.3E+07 0.57 Combo1 alum Staph19 2.3E+07 0.80 IsdBalum LAC 3.50E+07 2.67 Sta011 alum LAC 3.50E+07 1.35 EsxAB alum LAC3.50E+07 2.21 HlaH35L alum LAC 3.50E+07 0.71 Sta006 alum LAC 3.50E+072.39 Combo1 alum LAC 3.50E+07 2.66 IsdB alum MW2 3.00E+07 1.17 Sta011alum MW2 3.00E+07 0.82 EsxAB alum MW2 3.00E+07 1.39 HlaH35L alum MW23.00E+07 0.87 Sta006 alum MW2 3.00E+07 0.91 Combo1 alum MW2 3.00E+072.69 IsdB alum LAC 4.00E+07 1.54 Sta011 alum LAC 4.00E+07 1.95 EsxABalum LAC 4.00E+07 1.31 HlaH35L alum LAC 4.00E+07 0.75 Sta006 alum LAC4.00E+07 1.74 Combo1 alum LAC 4.00E+07 2.21 IsdB alum MW2 2.75E+07 1.22Sta011 alum MW2 2.75E+07 1.25 EsxAB alum MW2 2.75E+07 1.16 HlaH35L alumMW2 2.75E+07 1.61 Sta006 alum MW2 2.75E+07 1.13 Combo1 alum MW2 2.75E+071.97 Sta011 alum Mu-50 4.00E+07 1.10 EsxAB alum Mu-50 4.00E+07 0.86HlaH35L alum Mu-50 4.00E+07 0.71 Sta006 alum Mu-50 4.00E+07 1.57 Combo1alum Mu-50 4.00E+07 1.72 Sta011 alum Staph19 5.30E+07 1.23 EsxAB alumStaph19 5.30E+07 1.19 HlaH35L alum Staph19 5.30E+07 0.65 Sta006 alumStaph19 5.30E+07 2.00 Combo1 alum Staph19 5.30E+07 2.02 Sta011 alumMu-50 4.30E+07 1.33 EsxAB alum Mu-50 4.30E+07 0.36 HlaH35L alum Mu-504.30E+07 0.11 Sta006 alum Mu-50 4.30E+07 1.05 Combo1 alum Mu-50 4.30E+071.34 Sta011 alum Staph19 4.40E+07 1.07 EsxAB alum Staph19 4.40E+07 0.94HlaH35L alum Staph19 4.40E+07 1.19 Sta006 alum Staph19 4.40E+07 2.31Combo1 alum Staph19 4.40E+07 2.45 *alum = aluminium hydroxide **Logreduction in kidney CFU

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1. An immunogenic composition comprising a combination of antigens, saidcombination comprising two or more antigens selected from the groupconsisting of: (1) a sta006 antigen; (2) a sta011 antigen; (3) a esxAantigen; (4) a esxB antigen; (5) a hla antigen; (6) a ebpS antigen; (7)a efb antigen; (8) a emp antigen; (9) a esaC antigen; (10) a coAantigen; (11) a eap antigen; (12) a FnBA antigen; (13) a FnBB antigen;(14) a ebhA antigen; (15) a hlgB antigen; (16) a hlgC antigen; (17) aisdA antigen; (18) a isdB antigen; (19) a isdC antigen; (20) a isdGantigen; (21) a isdH antigen; (22) a isdI antigen; (23) a lukD antigen;(24) a lukE antigen; (25) a lukF antigen; (26) a lukS antigen; (27) anuc antigen; (28) a sasA antigen; (29) a sasB antigen; (30) a sasCantigen; (31) a sasD antigen; (32) a sasF antigen; (33) a sdrC antigen;(34) a sdrD antigen; (35) a sdrE2 antigen; (36) a spa antigen; (37) aclfA antigen; (38) a clfB antigen; (39) a sta001 antigen; (40) a sta002antigen; (41) a sta003 antigen; (42) a sta004 antigen; (43) a sta005antigen; (44) a sta007 antigen; (45) a sta008 antigen; (46) a sta009antigen; (47) a sta010 antigen; (48) a sta012 antigen; (49) a sta013antigen; (50) a sta014 antigen; (51) a sta015 antigen; (52) a sta016antigen; (53) a sta017 antigen; (54) a sta018 antigen; (55) a sta019antigen; (56) a sta020 antigen; (57) a sta021 antigen; (58) a sta022antigen; (59) a sta023 antigen; (60) a sta024 antigen; (61) a sta025antigen; (62) a sta026 antigen; (63) a sta027 antigen; (64) a sta028antigen; (65) a sta029 antigen; (66) a sta030 antigen; (67) a sta031antigen; (68) a sta032 antigen; (69) a sta033 antigen; (70) a sta034antigen; (71) a sta035 antigen; (72) a sta036 antigen; (73) a sta037antigen; (74) a sta038 antigen; (75) a sta039 antigen; (76) a sta040antigen; (77) a sta041 antigen; (78) a sta042 antigen; (79) a sta043antigen; (80) a sta044 antigen; (81) a sta045 antigen; (82) a sta046antigen; (83) a sta047 antigen; (84) a sta048 antigen; (85) a sta049antigen; (86) a sta050 antigen; (87) a sta051 antigen; (88) a sta052antigen; (89) a sta053 antigen; (90) a sta054 antigen; (91) a sta055antigen; (92) a sta056 antigen; (93) a sta057 antigen; (94) a sta058antigen; (95) a sta059 antigen; (96) a sta060 antigen; (97) a sta061antigen; (98) a sta062 antigen; (99) a sta063 antigen; (100) a sta064antigen; (101) a sta065 antigen; (102) a sta066 antigen; (103) a sta067antigen; (104) a sta068 antigen; (105) a sta069 antigen; (106) a sta070antigen; (107) a sta071 antigen; (108) a sta072 antigen; (109) a sta073antigen; (110) a sta074 antigen; (111) a sta075 antigen; (112) a sta076antigen; (113) a sta077 antigen; (114) a sta078 antigen; (115) a sta079antigen; (116) a sta080 antigen; (117) a sta082 antigen; (118) a sta083antigen; (119) a sta084 antigen; (120) a sta085 antigen; (121) a sta086antigen; (122) a sta087 antigen; (123) a sta088 antigen; (124) a sta089antigen; (125) a sta090 antigen; (126) a sta091 antigen; (127) a sta092antigen; (128) a sta093 antigen; (129) a sta094 antigen; (130) a sta095antigen; (131) a sta096 antigen; (132) a sta097 antigen; (133) a sta098antigen; (134) a sta099 antigen; (135) a sta100 antigen; (136) a sta101antigen; (137) a sta102 antigen; (138) a sta103 antigen; (139) a sta104antigen; (140) a sta105 antigen; (141) a sta106 antigen; (142) a sta107antigen; (143) a sta108 antigen; (144) a sta109 antigen; (145) a sta110antigen; (146) a sta111 antigen; (147) a sta112 antigen; (148) a sta113antigen; (149) a sta114 antigen; (150) a sta115 antigen; (151) a sta116antigen; (152) a sta117 antigen; (153) a sta118 antigen; (154) a sta119antigen; (155) a sta120 antigen; (156) a NW_(—)6 antigen; (157) aNW_(—)9 antigen; (158) a NW_(—)10 antigen; (159) a NW_(—)7 antigen;(160) a NW_(—)8 antigen; (161) a NW_(—)2 antigen; (162) a NW_(—)1antigen; (163) a sta081 antigen; and (164) a NW_(—)5 antigen.
 2. Thecomposition of claim 1, comprising at least one antigen selected fromnumbers (3) to (38) and at least one antigen selected from numbers (1),(2) and (37) to (149).
 3. The composition of claim 2, comprising: atleast one antigen selected from numbers (37), (38), (8), (9), (3), (4),(5), (17), (18), (19), (31), (32), (33), (34), (35) and (36); and atleast one antigen selected from (40), (1), (43), (2), (64), (96), (133)and (147).
 4. The composition of claim 1, comprising two or moreantigens selected from the group consisting of: (1) a clfA antigen; (2)a clfB antigen; (3) a sdrE2 antigen; (4) a sdrC antigen; (5) a SasFantigen; (6) a emp antigen; (7) a sdrD antigen; (8) a spa antigen; (9) aesaC antigen; (10) a esxA antigen; (11) a esxB antigen; (12) a sta006antigen; (13) a isdC antigen; (14) a hla antigen; (15) a sta011 antigen;(16) isdA antigen; (17) a isdB antigen; (18) a sasF antigen.
 5. Thecomposition of claim 1, comprising two or more antigens selected fromthe group consisting of: (1) a esxA antigen; (2) a esxB antigen; (3) asta006 antigen; (4) a hla antigen; (5) a sta011 antigen.
 6. Thecomposition of claim 1, comprising: (1) a esxA, antigen; (2) a esxBantigen; (3) a sta006 antigen; (4) a hla antigen; (5) a sta011 antigen.7. The composition of claim 5, wherein one or more of said antigens isadsorbed to an aluminium hydroxide adjuvant, and optionally wherein thecomposition includes a histidine buffer.
 8. The composition of claim 5,further comprising: one or more conjugates of (i) a S. aureusexopolysaccharide and (ii) a carrier protein.
 9. The composition ofclaim 5, further comprising: one or more conjugates of (i) a S. aureuscapsular polysaccharide and (ii) a carrier protein.
 10. A polypeptide offormula NH₂-A-{-X-L-}_(n)-B—COOH, wherein: X is an amino acid sequenceof a staphylococcal antigen, selected from the group consisting of S.aureus antigens sta006, sta011, esxA, esxB, hla, clfA, clfB, coA, eap,ebhA, ebpS, efb, emp, esaC, FnBA, FnBB, hlgB, hgC, isdA, isdB, isdC,isdG, isdH, isdI, lukD, lukE, lukF, lukS, nuc, sasA, sasB, sasC, sasD,sasF, sdrC, sdrD, sdrE2, spa, sta001, sta002, sta003, sta004, sta005,sta007, sta008, sta009, sta010, sta012, sta013, sta014, sta015, sta016,sta017, sta018, sta019, sta020, sta021, sta022, sta023, sta024, sta025,sta026, sta027, sta028, sta029, sta030, sta031, sta032, sta033, sta034,sta035, sta036, sta037, sta038, sta039, sta040, sta041, sta042, sta043,sta044, sta045, sta046, sta047, sta048, sta049, sta050, sta051, sta052,sta053, sta054, sta055, sta056, sta057, sta058, sta059, sta060, sta061,sta062, sta063, sta064, sta065, sta066, sta067, sta068, sta069, sta070,sta071, sta072, sta073, sta074, sta075, sta076, sta077, sta078, sta079,sta080, sta081, sta082, sta083, sta084, sta085, sta086, sta087, sta088,sta089, sta090, sta091, sta092, sta093, sta094, sta095, sta096, sta097,sta098, sta099, sta100, sta101, sta102, sta103, sta104, sta105, sta106,sta107, sta108, sta109, sta110, sta111, sta112, sta113, sta114, sta115,sta116, sta117, sta118, NW_(—)6, NW_(—)9, NW_(—)10, NW_(—)7, NW_(—)8,NW_(—)2, NW_(—)1, and NW_(—)5; L is an optional linker amino acidsequence; A is an optional N-terminal amino acid sequence; B is anoptional C-terminal amino acid sequence; and n is an integer of 2 ormore.
 11. An immunogenic composition comprising the polypeptide of claim10 and further comprising: (A) one or more conjugates of (i) a S. aureusexopolysaccharide and (ii) a carrier protein; and/or (B) one or moreconjugates of (i) a S. aureus capsular polysaccharide and (ii) a carrierprotein.
 12. The composition of claim 4, wherein the clfA antigen canelicit an antibody which recognises SEQ ID NO: 1 and comprises an aminoacid sequence; (a) having 80% or more identity to SEQ ID NO: 1; and/or(b) comprising a fragment of at least 7 consecutive amino acids of SEQID NO: 1, wherein the fragment comprises an epitope from SEQ ID NO: 1.13. A polypeptide comprising an amino acid sequence (a) having 80% ormore identity to SEQ ID NO: 151; and/or (b) comprising a fragment of atleast 7 consecutive amino acids from amino acids 1-97 of SEQ ID NO: 151and at least 7 consecutive amino acids from amino acids 104-207 of SEQID NO: 151, wherein the polypeptide can elicit antibodies whichrecognise both the wild-type staphylococcal protein comprising SEQ IDNO: 10 and the wild-type staphylococcal protein comprising SEQ ID NO:11.
 14. An immunogenic composition comprising the polypeptide of claim13 and one or more of (i) a sta006 antigen; (ii) a hla antigen; and/or(iii) a sta011 antigen.
 15. An immunogenic composition comprising thepolypeptide of claim 13 and (i) a sta006 antigen; (ii) a hla antigen;and (iii) a sta011 antigen.
 16. The composition of claim 14, includingan adjuvant.
 17. A polypeptide comprising amino acid sequence having 80%or more identity to an amino acid sequence selected from SEQ ID NOs:151, 152, 168, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,220, 221, 222, 223, 224, 237, 238,
 241. 18. A polypeptide comprising anamino acid sequence selected from SEQ ID NOs: 151, 152, 168, 202, 203,204, 205, 206, 207, 208, 209, 210, 211, 212, 220, 221, 222, 223, 224,237, 238, 241, 242, 243, 244,
 245. 19. A polypeptide comprising: (a) afirst sequence having 90% or more identity to SEQ ID NO: 218; and (b) asecond sequence having 90% or more identity to SEQ ID NO: 219, whereinthe first and second sequences are either directly joined or are joinedby an intervening amino acid sequence having fewer than 10 amino acids.20. A pharmaceutical composition comprising the polypeptide of claim 17,21. A method for raising an immune response in a mammal comprising thestep of administering to the mammal an effective amount of thepolypeptide or composition of claim
 5. 22. Nucleic acid encoding thepolypeptide of claim 17.